Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment

ABSTRACT

The present invention encompasses novel antibodies and fragments thereof which immunospecifically bind to one or more RSV antigens and compositions comprising said antibodies and antibody fragments. The present invention encompasses methods preventing respiratory syncytial virus (RSV) infection in a human, comprising administering to said human a prophylactically effective amount of one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens, wherein a certain serum titer of said antibodies or antibody fragments is achieved in said human subject. The present invention also encompasses methods for treating or ameliorating symptoms associated with a RSV infection in a human, comprising administering to said human a therapeutically effective amount of one or more antibodies or fragments thereof that immunospecifically bind to one or more RSV antigens, wherein a certain serum titer of said antibodies or antibody fragments is achieved in said human subject. The present invention further encompasses compositions comprising antibodies or fragments thereof that immunospecifically bind to a RSV antigen, and methods using said compositions for detection or diagnosis a RSV infection

[0001] This application is a continuation-in-part of U.S. applicationSer. No. 09/724,396, filed Nov. 28, 2001, the entire contents of whichis incorporated herein by reference.

1. INTRODUCTION

[0002] The present invention relates to compositions comprisingantibodies or fragments thereof that immunospecifically bind to a RSVantigen and methods for preventing, treating or ameliorating symptomsassociated with respiratory syncytial virus (RSV) infection utilizingsaid compositions. In particular, the present invention relates tomethods for preventing, treating or ameliorating symptoms associatedwith RSV infection, said methods comprising administering to a humansubject an effective amount of one or more antibodies or fragmentsthereof that immunospecifically bind to a RSV antigen, wherein a certainserum titer of said antibodies or antibody fragments is achieved in saidhuman subject. The present invention also relates to detectable ordiagnostic compositions comprising antibodies or fragments thereof thatimmunospecifically bind to a RSV antigen and methods for detecting ordiagnosing RSV infection utilizing said compositions.

2. BACKGROUND OF THE INVENTION

[0003] Respiratory syncytial virus (RSV) is the leading cause of seriouslower respiratory tract disease in infants and children (Feigen et al.,eds., 1987, In: Textbook of Pediatric Infectious Diseases, W B Saunders,Philadelphia at pages 1653-1675; New Vaccine Development, EstablishingPriorities, Vol. 1, 1985, National Academy Press, Washington D.C. atpages 397-409; and Ruuskanen et al., 1993, Curr. Probl. Pediatr.23:50-79). The yearly epidemic nature of RSV infection is evidentworldwide, but the incidence and severity of RSV disease in a givenseason vary by region (Hall, C. B., 1993, Contemp. Pediatr. 10:92- 110).In temperate regions of the northern hemisphere, it usually begins inlate fall and ends in late spring. Primary RSV infection occurs mostoften in children from 6 weeks to 2 years of age and uncommonly in thefirst 4 weeks of life during nosocomial epidemics (Hall et al., 1979,New Engl. J. Med. 300:393-396). Children at increased risk from RSVinfection include preterm infants (Hall et al., 1979, New Engl. J. Med.300:393-396) and children with bronchopulmonary dysplasia (Groothuis etal., 1988, Pediatrics 82:199-203), congenital heart disease (MacDonaldet al., New Engl. J. Med. 307:397-400), congenital or acquiredimmunodeficiency (Ogra et al., 1988, Pediatr. Infect. Dis. J. 7:246-249;and Pohl et al., 1992, J. Infect. Dis. 165:166-169), and cystic fibrosis(Abman et al., 1988, J. Pediatr. 113:826-830). The fatality rate ininfants with heart or lung disease who are hospitalized with RSVinfection is 3%-4% (Navas et al., 1992, J. Pediatr. 121:348-354).

[0004] RSV infects adults as well as infants and children. In healthyadults, RSV causes predominantly upper respiratory tract disease. It hasrecently become evident that some adults, especially the elderly, havesymptomatic RSV infections more frequently than had been previouslyreported (Evans, A. S., eds., 1989, Viral Infections of Humans.Epidemiology and Control, 3^(rd) ed., Plenum Medical Book, New York atpages 525-544). Several epidemics also have been reported among nursinghome patients and institutionalized young adults (Falsey, A. R., 1991,Infect. Control Hosp. Epidemiol. 12:602-608; and Garvie et al., 1980,Br. Med. J. 281:1253-1254). Finally, RSV may cause serious disease inimmunosuppressed persons, particularly bone marrow transplant patients(Hertz et al., 1989, Medicine 68:269-281).

[0005] Treatment options for established RSV disease are limited. SevereRSV disease of the lower respiratory tract often requires considerablesupportive care, including administration of humidified oxygen andrespiratory assistance (Fields et al., eds, 1990, Fields Virology,2^(nd) ed., Vol. 1, Raven Press, New York at pages 1045-1072). The onlydrug approved for treatment of infection is the antiviral agentribavirin (American Academy of Pediatrics Committee on InfectiousDiseases, 1993, Pediatrics 92:501-504). It has been shown to beeffective in the treatment of RSV pneumonia and bronchiolitis, modifyingthe course of severe RSV disease in immunocompetent children (Smith etal., 1991, New Engl. J. Med. 325:24-29). However, ribavirin has hadlimited use because it requires prolonged aerosol administration andbecause of concerns about its potential risk to pregnant women who maybe exposed to the drug during its administration in hospital settings.

[0006] While a vaccine might prevent RSV infection, no vaccine is yetlicensed for this indication. A major obstacle to vaccine development issafety. A formalin-inactivated vaccine, though immunogenic, unexpectedlycaused a higher and more severe incidence of lower respiratory tractdisease due to RSV in immunized infants than in infants immunized with asimilarly prepared trivalent parainfluenza vaccine (Kim et al., 1969,Am. J. Epidemiol. 89:422-434; and Kapikian et al., 1969, Am. J.Epidemiol. 89:405-421). Several candidate RSV vaccines have beenabandoned and others are under development (Murphy et al., 1994, VirusRes. 32:13-36), but even if safety issues are resolved, vaccine efficacymust also be improved. A number of problems remain to be solved.Immunization would be required in the immediate neonatal period sincethe peak incidence of lower respiratory tract disease occurs at 2-5months of age. The immaturity of the neonatal immune response togetherwith high titers of maternally acquired RSV antibody may be expected toreduce vaccine immunogenicity in the neonatal period (Murphy et al.,1988, J. Virol. 62:3907-3910; and Murphy et al., 1991, Vaccine9:185-189). Finally, primary RSV infection and disease do not protectwell against subsequent RSV disease (Henderson et al., 1979, New Engl.J. Med. 300:530-534).

[0007] Currently, the only approved approach to prophylaxis of RSVdisease is passive immunization. Initial evidence suggesting aprotective role for IgG was obtained from observations involvingmaternal antibody in ferrets (Prince, G. A., Ph.D. diss., University ofCalifornia, Los Angeles, 1975) and humans (Lambrecht et al, 1976, J.Infect. Dis. 134:211-217; and Glezen et al., 1981, J. Pediatr.98:708-715). Hemming et al. (More et al., eds., 1986, Clinical Use ofIntravenous Immunoglobulins, Academic Press, London at pages 285-294)recognized the possible utility of RSV antibody in treatment orprevention of RSV infection during studies involving thepharmacokinetics of an intravenous immune globulin (IVIG) in newbornssuspected of having neonatal sepsis. They noted that 1 infant, whoserespiratory secretions yielded RSV, recovered rapidly after IVIGinfusion. Subsequent analysis of the IVIG lot revealed an unusually hightiter of RSV neutralizing antibody. This same group of investigatorsthen examined the ability of hyperimmune serum or immune globulin,enriched for RSV neutralizing antibody, to protect cotton rats andprimates against RSV infection (Prince et al., 1985, Virus Res.3:193-206; Prince et al., 1990, J. Virol. 64:3091-3092; Hemming et al.,1985, J. Infect. Dis. 152:1083-1087; Prince et al., 1983, Infect. Immun.42:81-87; and Prince et al., 1985, J. Virol. 55:517-520). Results ofthese studies suggested that RSV neutralizing antibody givenprophylactically inhibited respiratory tract replication of RSV incotton rats. When given therapeutically, RSV antibody reduced pulmonaryviral replication both in cotton rats and in a nonhuman primate model.Furthermore, passive infusion of immune serum or immune globulin did notproduce enhanced pulmonary pathology in cotton rats subsequentlychallenged with RSV.

[0008] Recent clinical studies have demonstrated the ability of thispassively administered RSV hyperimmune globulin (RSV IVIG) to protectat-risk children from severe lower respiratory infection by RSV(Groothius et al., 1993, New Engl. J. Med. 329:1524-1530; and ThePREVENT Study Group, 1997, Pediatrics 99:93-99). While this is a majoradvance in preventing RSV infection, this treatment poses certainlimitations in its widespread use. First, RSV IVIG must be infusedintravenously over several hours to achieve an effective dose. Second,the concentrations of active material in hyperimmune globulins areinsufficient to treat adults at risk or most children with comprisedcardiopulmonary function. Third, intravenous infusion necessitatesmonthly hospital visits during the RSV season. Finally, it may provedifficult to select sufficient donors to produce a hyperimmune globulinfor RSV to meet the demand for this product. Currently, onlyapproximately 8% of normal donors have RSV neutralizing antibody titershigh enough to qualify for the production of hyperimmune globulin.

[0009] One way to improve the specific activity of the immunoglobulinwould be to develop one or more highly potent RSV neutralizingmonoclonal antibodies (MAbs). Such MAbs should be human or humanized inorder to retain favorable pharmacokinetics and to avoid generating ahuman anti-mouse antibody response, as repeat dosing would be requiredthroughout the RSV season. Two glycoproteins, F and G, on the surface ofRSV have been shown to be targets of neutralizing antibodies (Fields etal., 1990, supra; and Murphy et al., 1994 , supra). These two proteinsare also primarily responsible for viral recognition and entry intotarget cells; G protein binds to a specific cellular receptor and the Fprotein promotes fusion of the virus with the cell. The F protein isalso expressed on the surface of infected cells and is responsible forsubsequent fusion with other cells leading to syncytia formation. Thus,antibodies to the F protein may directly neutralize virus or block entryof the virus into the cell or prevent syncytia formation. Althoughantigenic and structural differences between A and B subtypes have beendescribed for both the G and F proteins, the more significant antigenicdifferences reside on the G glycoprotein, where amino acid sequences areonly 53% homologous and antigenic relatedness is 5% (Walsh et al., 1987,J.

[0010] Infect. Dis. 155:1198-1204; and Johnson et al., 1987, Proc. Natl.Acad. Sci. USA 84:5625-5629). Conversely, antibodies raised to the Fprotein show a high degree of cross-reactivity among subtype A and Bviruses. Beeler and Coelingh (1989, J. Virol. 7:2941-2950) conducted anextensive analysis of 18 different murine MAbs directed to the RSV Fprotein. Comparison of the biologic and biochemical properties of theseMAbs resulted in the identification of three distinct antigenic sites(designated A, B, and C). Neutralization studies were performed againsta panel of RSV strains isolated from 1956 to 1985 that demonstrated thatepitopes within antigenic sites A and C are highly conserved, while theepitopes of antigenic site B are variable.

[0011] A humanized antibody directed to an epitope in the A antigenicsite of the F protein of RSV, SYNAGIS®, is approved for intramuscularadministration to pediatric patients for prevention of serious lowerrespiratory tract disease caused by RSV at recommended monthly doses of15 mg/kg of body weight throughout the RSV season (November throughApril in the northern hemisphere). SYNAGIS® is a composite of human(95%) and murine (5%) antibody sequences. See, Johnson et al., 1997, J.Infect. Diseases 176:1215-1224 and U.S. Pat. No. 5,824,307, the entirecontents of which are incorporated herein by reference. The human heavychain sequence was derived from the constant domains of human IgG₁ andthe variable framework regions of the VH genes or Cor (Press et al.,1970, Biochem. J. 117:641-660) and Cess (Takashi et al., 1984, Proc.Natl. Acad. Sci. USA 81:194-198). The human light chain sequence wasderived from the constant domain of Cκ and the variable frameworkregions of the VL gene K104 with Jκ-4 (Bentley et al., 1980, Nature288:5194-5198). The murine sequences derived from a murine monoclonalantibody, Mab 1129 (Beeler et al., 1989, J. Virology 63:2941-2950), in aprocess which involved the grafting of the murine complementaritydetermining regions into the human antibody frameworks.

[0012] Although SYNAGIS® has been successfully used for the preventionof RSV infection in pediatric patients, multiple intramuscular doses of15 mg/kg of SYNAGIS™ is required to achieve a prophylactic effect. Inpediatric patients less than 24 months of age, the mean half-life ofSYNAGIS® has been shown to be 20 days and monthly intramuscular doses of15 mg/kg have been shown to result in a mean ± standard derivation 30day serum titer of 37±21 μg/ml after the first injection, 57±41 μg/mlafter the second injection, 68±51 μg/ml after the third injection, and72±50 μg/ml after the fourth injection (The IMpact RSV Study Group,1998, Pediatrics 102:531-537). Serum concentrations of greater than 30μg/ml have been shown to be necessary to reduce pulmonary RSVreplication by 100 fold in the cotton rat model of RSV infection.However, the administration of multiple intramuscular doses of 15 mg /kgof antibody is inconvenient for the patient. Thus, a need exists forantibodies that immunospecifically bind to a RSV antigen, which arehighly potent, have an improved pharmacokinetic profile, and thus havean overall improved therapeutic profile. Further, a need exists forantibodies that immunospecifically bind to a RSV antigen which requireless frequent administration.

[0013] Citation or discussion of a reference herein shall not beconstrued as an admission that such is prior art to the presentinvention.

3. SUMMARY OF THE INVENTION

[0014] The present invention is based, in part, on the development ofmethods for achieving or inducing a prophylactically or therapeuticallyeffective serum titer of an antibody or fragment thereof thatimmunospecifically binds to a respiratory syncytial virus (RSV) antigenin a mammal by passive immunization with such an antibody or fragmentthereof, which methods require lower dosages and/or less frequentadministration than previously known methods. The present invention isalso based, in part, on the identification of antibodies with higheraffinities for a RSV antigen which result in increased efficacy forprophylactic or therapeutic uses such that lower serum titers areprophylactically or therapeutically effective, thereby permittingadministration of lower dosages and/or reduced frequency ofadministration.

[0015] The present invention provides methods of preventing,neutralizing, treating and ameliorating one or more symptoms associatedwith RSV infection in a subject comprising administering to said subjectone or more antibodies or fragments thereof which immunospecificallybind to one or more RSV antigens with high affinity and/or high avidity.Because a lower serum titer of such antibodies or antibody fragments istherapeutically or prophylactically effective than the effective serumtiter of known antibodies, lower doses of said antibodies or antibodyfragments can be used to achieve a serum titer effective for theprevention, neutralization, treatment and the amelioration of symptomsassociated with a RSV infection. The use of lower doses of antibodies orfragments thereof which immunospecifically bind to one or more RSVantigens reduces the likelihood of adverse effects. Further, the highaffinity and/or high avidity of the antibodies of the invention orfragments thereof enable less frequent administration of said antibodiesor antibody fragments than previously thought to be necessary for theprevention, neutralization, treatment or the amelioration of symptomsassociated with a RSV infection.

[0016] The present invention also provides antibodies whichimmunospecifically bind to one or more RSV antigens and have increasedin vivo half-lives relative to known antibodies such as, e.g., SYNAGIS®.In particular, the present invention encompasses antibodies whichimmunospecifically bind to one or more RSV antigens and have increasedin vivo half-lives relative to known antibodies (e.g., SYNAGIS®), saidincreased half-lives resulting from one or more modifications (e.g.,substitutions, deletions, or insertions) in amino acid residuesidentified to be involved in the interaction of the Fc domain of saidantibodies and the FcRn receptor. The present invention also encompassespegylated antibodies and fragments thereof which immunospecifically bindto one or more RSV antigens and have increased in vivo half-livesrelative to known antibodies such as, e.g., SYNAGIS®. The increased invivo half-lives of antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens reduce the dosageand/or frequency of administration of said antibodies or fragmentsthereof to a subject.

[0017] The invention encompasses sustained release formulations for theadministration of one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens to a subject. Thesustained release formulations reduce the dosage and/or frequency ofadministration of said antibodies or antibody fragments to a subject.Further, the sustained release formulations may be administered tomaintain a therapeutically or prophylactically effective serum titerwhich does not exceed a certain maximum serum titer for a certain periodof time.

[0018] The present invention encompasses methods of delivering one ormore antibodies or fragments thereof which immunospecifically bind toone or more RSV antigens directly to the site of RSV infection. Inparticular, the invention encompasses pulmonary delivery of one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens. The improved methods of delivering of one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens reduce the dosage and/or frequency of administrationof said antibodies or antibody fragments to a subject.

[0019] The present invention provides antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens and have anassociation rate constant or k_(on) rate (antibody (AB)+antigen

[0020] of at least 10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹. In particular, the present inventionprovides compositions for use in the prevention, treatment oramelioration of one or more symptoms associated with a RSV infection,said compositions comprising one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens and have an ak_(on) rate of at least 10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹.

[0021] The present invention provides antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens and have ak_(off) rate (antibody (AB)+antigen

[0022] of less than 10⁻¹ s⁻¹, less than 5×10⁻¹ s⁻¹, less than 10⁻² s⁻¹,less than 5×10⁻² s⁻¹, less than 10⁻³ s⁻¹, less than 5×10⁻³ s⁻¹, lessthan 10⁻⁴ s⁻¹, less than 5×10⁻⁴ s⁻¹, less than 10⁻⁵ s⁻¹, less than5×10⁻⁵ s⁻¹, less than 10⁻⁶ s⁻¹, less than 5×10⁻⁶ s⁻¹, less than 10⁻⁷s⁻¹, less than 5×10⁻⁷ s⁻¹, less than 10⁻⁸ s⁻¹, less than 5×10⁻⁸ s⁻¹,less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹ s⁻¹, or less than 10⁻¹⁰ s⁻¹. Inparticular, the present invention provides compositions for use in theprevention, treatment or amelioration of one or more symptoms associatedwith a RSV infection, said compositions comprising one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens and have a k_(off) rate of less than 10⁻¹ s⁻¹, lessthan 5×10⁻¹ s⁻¹, less than 10⁻² s⁻¹, less than 5×10⁻² s⁻¹, less than10³¹ ³s⁻¹, less than 5×10⁻³ s⁻¹, less than 10⁻⁴ s⁻¹, less than 5×10⁻⁴s⁻¹, less than 10⁻⁵ s⁻¹, less than 5×10⁻⁵ S⁻¹, less than 10⁻⁶ s⁻¹, lessthan 5×10⁻⁶ s⁻¹, less than 10⁻⁷ s⁻¹, less than 5×10⁻⁷ s⁻¹, less than10⁻⁸ s⁻¹, less than 5×10⁻⁸ s⁻¹, less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹s⁻¹, or less than 10⁻¹⁰ s⁻¹.

[0023] The present invention also provides antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens andhave an affinity constant or K_(a) (k_(on)/k_(off)) of at least 10² M⁻¹,at least 5×10² M⁻¹, at least 10³ M⁻¹, at least 5×10³ M⁻¹, at least 10⁴M⁻¹, at least 5×10⁴ M⁻¹, at least 10⁵ M⁻¹, at least 5×10⁵ M⁻¹, at least10⁶ M⁻¹, at least 5×10⁶ M⁻¹, at least 10⁷ M⁻¹, at least 5×10⁷M⁻¹, atleast 10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹,at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹,at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹. In particular, the present inventionprovides compositions for use in the prevention, treatment oramelioration of one or more symptoms associated with a RSV infection,said compositions comprising one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens and have aK_(a) of at least 10² M⁻¹, at least 5×10² M⁻¹, at least 10³ M⁻¹, atleast 5×10³ M⁻¹, at least 10⁴ M⁻¹, at least 5×10⁴ M⁻¹, at least 10⁵ M⁻¹,at least 5×10⁵ M⁻¹, at least 10⁶ M⁻¹, at least 5×10⁶ M⁻¹, at least 10⁷M⁻¹, at least 5×10⁷M⁻¹, at least 10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, atleast 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹²M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least5×10¹⁴ M⁻¹ at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹.

[0024] The present invention provides antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens and have amedian effective concentration (EC₅₀) of less than 0.01 nM, less than0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, lessthan 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, lessthan 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitromicroneutralization assay In particular, the present invention providescompositions for use in the prevention, treatment or amelioration of oneor more symptoms associated with a RSV infection, said compositionscomprising one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens and have an EC₅₀ ofless than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than2 nM, in an in vitro microneutralization assay

[0025] The present invention also provides antibodies or fragmentsthereof comprising a VH domain having the amino acid sequence of any VHdomain listed in Table 2 and compositions comprising said antibodies orantibody fragments for use in the prevention, treatment or ameliorationof one or more symptoms associated with a RSV infection. The presentinvention also provides antibodies or fragments thereof comprising oneor more VH complementarity determining regions (CDRs) having the aminoacid sequence of one or more VH CDRs listed in Table 2 and/or Table 3and compositions comprising said antibodies or antibody fragments foruse in the prevention, treatment or amelioration of one or more symptomsassociated with a RSV infection. The present invention also providesantibodies or fragments thereof comprising a VL domain having the aminoacid sequence of any VL domain listed in Table 2. The present inventionalso provides antibodies or fragments thereof comprising one or more VLCDRs having the amino acid sequence of one or more VL CDRs listed inTable 2 and/or Table 3 and compositions comprising said antibodies orantibody fragments for use in the prevention, treatment or ameliorationof one or more symptoms associated with a RSV infection. The presentinvention further provides antibodies comprising a VH domain and a VLdomain having the amino acid sequence of any VH domain and VL domainlisted in Table 2 and compositions comprising said antibodies orantibody fragments for use in the prevention, treatment or ameliorationof one or more symptoms associated with a RSV infection. The presentinvention further provides antibodies comprising one or more VH CDRs andone or more VL CDRs having the amino acid sequence of one or more VHCDRs and one or more VL CDRs listed in Table 2 and/or 3 and compositionscomprising said antibodies or antibody fragments for use in theprevention, treatment or amelioration of one or more symptoms associatedwith a RSV infection. In the above embodiments, preferably the antibodybinds immunospecifically to a RSV antigen.

[0026] The present invention also encompasses methods for achieving aserum titer of at least 40 μg/ml of one or more antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens in amammal, preferably a primate and most preferably a human. In particular,the present invention provides methods for achieving a serum titer of atleast 40 μg/ml (preferably at least 75 μg/ml, more preferably at least100 μg/ml, and most preferably at least 150 μg/ml) of an antibody orfragment thereof that immunospecifically binds to a RSV antigen in anon-primate mammal, comprising administering a dose of less than 2.5mg/kg (preferably 1.5 mg/kg or less) of the antibody or antibodyfragment to the non-primate mammal and measuring the serum titer of theantibody or antibody fragment at least 1 day after administering thedose to the non-primate mammal. The present invention also providesmethods for achieving a serum titer of at least 150 μg/ml (preferably atleast 200 μg/ml) of an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen in a non-primate mammal,comprising administering a dose of approximately 5 mg/kg of the antibodyor antibody fragment to the non-primate mammal and measuring the serumtiter of the antibody or antibody fragment at least I day after theadministration of the dose to the non-primate mammal.

[0027] The present invention also provides methods for achieving a serumtiter of at least 40 μg/ml of an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen in a primate, comprisingadministering a first dose of 10 mg/kg (preferably 5 mg/kg or less andmore preferably 1.5 mg/kg or less) of the antibody or antibody fragmentto the primate and measuring the serum titer of the antibody or antibodyfragment 20 days (preferably 25, 30, 35 or 40 days) after administratingthe first dose to the primate and prior to the administration of anysubsequent dose. The present invention also provides methods forachieving a serum titer of at least 75 μg/ml (preferably at least 100μg/ml, at least 150 μg/ml, or at least 200 μg/ml) of an antibody orfragment thereof that immunospecifically binds to a RSV antigen in aprimate, comprising administering a first dose of approximately 15 mg/kgof the antibody or antibody fragment to the primate and measuring theserum titer of the antibody or antibody fragment 20 days (preferably 25,30, 35 or 40 days) after administering the first dose to the primate butprior to any subsequent dose.

[0028] The present invention also provides methods for preventing,treating, or ameliorating one or more symptoms associated with a RSVinfection in a human subject, said methods comprising administering tosaid human subject at least a first dose of approximately 15 mg/kg of anantibody or fragment thereof that immunospecifically binds to a RSVantigen so that said human subject has a serum antibody titer of atleast 75 μg/ml, preferably at least 100 μg/ml, at least 150 μg/ml, or atleast 200 μg/ml 30 days after the administration of the first dose ofthe antibody or antibody fragment and prior to the administration of asubsequent dose. The present invention also provides methods forpreventing, treating or ameliorating one or more symptoms associatedwith a RSV infection in a human subject, said methods comprisingadministering to said human subject at least a first dose of less than15 mg/kg (preferably 10 mg/kg or less, more preferably 5 mg/kg or less,and most preferably 1.5 mg/kg or less) of an antibody or fragmentthereof that immunospecifically binds to a RSV antigen so that saidhuman subject has a serum antibody titer of at least 75 μg/ml,preferably at least 100 μg/ml, at least 150 μg/ml, or at least 200 μg/ml30 days after the administration of the first dose of the antibody orantibody fragment and prior to the administration of a subsequent dose.The present invention further provides methods for preventing, treatingor ameliorating one or more symptoms associated with a RSV infection ina human subject, said methods comprising administering to said humansubject a first dose of an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen such that a prophylacticallyor therapeutically effective serum titer of less than 10 μg/ml isachieved no more than 30 days after administering the antibody orantibody fragment.

[0029] The present invention provides methods for achieving atherapeutically or prophylactically effective serum titer in a mammal,said methods comprising administering to said mammal an antibody orfragment thereof which immunospecifically binds to a RSV antigen andwhich has a

[0030] of at least 2.5×10⁻⁵ M⁻¹ s′, preferably at least 3×10⁵M⁻¹ s⁻¹, atleast 5×10⁵M⁻¹ s⁻¹, at least 10⁶M⁻¹ s⁻¹, at least 5×10⁶M⁻¹ s⁻¹, at least10⁷M⁻¹ s⁻¹, at least 5×10⁷M⁻¹ s⁻¹ or at least 10⁸M⁻¹ s⁻¹. In particular,the present invention provides methods for achieving a therapeuticallyor prophylactically effective serum titer, wherein said effective serumtiter is less than 30 μg/ml (and is preferably at least 2 μg/ml, morepreferably at least 4 μg/ml, and most preferably at least 6 μg/ml) aftera certain number of days (for example, but not limited to, 20, 25, 30 or35 days) without any other dosing within that period, comprisingadministering to a mammal an antibody or fragment thereof whichimmunospecifically binds to a RSV antigen and which has a k_(on) rate ofat least 2.5×10⁵ M⁻¹s⁻¹, preferably at least 3×10⁵ M⁻¹s⁻¹, at least5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷M⁻¹s⁻¹, at least 5×10⁷ M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹. Preferably, theantibody or antibody fragment has a higher k₀, rate than SYNAGIS®.

[0031] The present invention also provides methods of neutralizing RSVusing an antibody or fragment thereof which immunospecifically bind to aRSV antigen and which has a k_(on) rate of at least 2.5×10⁵ M⁻¹s⁻¹,preferably at least 3×10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹ to achieve a prophylactically ortherapeutically effective serum titer, wherein said effective serumtiter is less than 30 μg/ml (and is preferably at least 2 μg/ml, morepreferably at least 4 μg/ml, and most preferably at least 6 μg/ml) 20,25, 30, or 35 days after administration without any other dosageadministration. Preferably, the antibody or antibody fragment has ahigher k_(on) rate than SYNAGIS®.

[0032] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, said methods comprisingadministering to said mammal, a dose of less than 15 mg/kg (preferably 5mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5mg/kg or less) of an antibody or fragment thereof whichimmunospecifically binds to a RSV antigen and has a k_(on) rate of atleast 2.5×10⁵ M⁻¹s⁻¹, preferably at least 3×10⁵ M⁻¹s⁻¹, at least 5×10⁵M⁻¹s⁻¹, at least 10⁶ M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹,at least 5×10⁷ M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹. Preferably, the antibodyor antibody fragment has a higher k_(on) rate for the RSV F glycoproteinthan SYNAGIS®.

[0033] The present invention also provides methods for achieving atherapeutically or prophylactically effective serum titer in a mammal,said methods comprising administering to said mammal an antibody orfragment thereof which immunospecifically binds to a RSV antigen andwhich has a

[0034] of less than 6.5×10⁻⁴ sec⁻¹, less than 5×10⁻⁴ sec⁻¹, less than3×10⁻⁴ sec⁻¹, less than 2×10⁻⁴ sec⁻¹, less than 1×10⁻⁴ sec⁻¹, or lessthan 5×10⁻³ sec⁻¹. In particular, the present invention provides methodsfor achieving a therapeutically or prophylactically effective serumtiter, wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) after a certain number of days (forexample, but not limited to, 20, 25, 30 or 35 days) without any otherdosing within that period, comprising administering to a mammal anantibody or fragment thereof which immunospecifically binds to a RSVantigen and which has a K_(off) rate of less than 6.5×10⁻⁴ sec⁻¹, lessthan 5×10⁻⁴ sec⁻¹, less than 3×10⁻⁴ sec⁻¹, less than 2×10⁻⁴ sec⁻¹, lessthan 1×10⁻⁴ sec⁻¹, or less than 3×10⁻³ sec⁻¹. Preferably, the antibodyor fragment thereof has a lower K_(off) rate than SYNAGIS®.

[0035] The present invention also provides methods of neutralizing RSVusing an antibody or antibody fragment thereof which immunospecificallybinds to a RSV antigen and which has a K_(off) rate of less than6.5×10⁻⁴ sec⁻¹, less than 5×10⁻⁴ sec⁻¹, less than 3×10⁻⁴ sec⁻¹, lessthan 2×10⁻⁴ sec⁻¹, less than 1×10⁴ sec⁻¹, or less than 5×10⁻¹ sec⁻¹ toachieve a prophylactically or therapeutically effective serum titer,wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) 20, 25, 30, or 35 days after administrationwithout any other dosage administration. Preferably, the antibody orantibody fragment has a lower K_(off) than SYNAGIS®.

[0036] The present invention also provides methods for preventing,treating, or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, said methods comprisingadministering to a said mammal a dose of less than 15 mg/kg (preferably5 mg/kg or less, more preferably 3 mg/kg or less, and most preferably1.5 mg/kg or less) of an antibody or a fragment thereof whichimmunospecifically binds to a RSV antigen and which has a K_(off) rateof less than 6.5×10⁻⁴ sec⁻¹, less than 5×10⁻⁴ sec⁻¹, less than 3×10⁻⁴sec⁻¹, less than 2×10⁻⁴ sec⁻¹, less than 1×10⁻⁴ sec⁻¹, or less than5×10⁻³ sec⁻¹. Preferably, the antibody or antibody fragment has a lowerK_(off) rate than SYNAGIS®.

[0037] The present invention also provides methods for achieving atherapeutically or prophylactically effective serum titer in a mammal,said methods comprising administering to said mammal an antibody orfragment thereof which immunospecifically binds to a RSV antigen andwhich has an EC₅₀ of less than 0.01 nM, less than 0.025 nM, less than0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5 nM, lessthan 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5 nM, lessthan 1.75 nM, or less than 2 nM, in an in vitro microneutralizationassay. In particular, the present invention provides methods forachieving a therapeutically or prophylactically effective serum titer,wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) after a certain number of days (forexample, but not limited to, 20, 25, 30 or 35 days) without any otherdosing within that period, comprising administering to a mammal anantibody or fragment thereof which immunospecifically binds to a RSVantigen and which has an EC₅₀ of less than 0.01 nM, less than 0.025 nM,less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, less than 0.5nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, less than 1.5nM, less than 1.75 nM, or less than 2 nM, in an in vitromicroneutralization assay Preferably, the antibody or antibody fragmenthas a lower EC₅₀ than SYNAGIS®.

[0038] The present invention also provides methods of neutralizing RSVusing an antibody or fragment thereof which immunospecifically binds toa RSV antigen and which has an EC₅₀ of less than 0.01 nM, less than0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25 nM, lessthan 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25 nM, lessthan 1.5 nM, less than 1.75 nM, or less than 2 nM, in an in vitromicroneutralization assay to achieve a prophylactically ortherapeutically effective serum titer, wherein said effective serumtiter is less than 30 μg/ml (and is preferably at least 2 μg/ml, morepreferably at least 4 μg/ml, and most preferably at least 6 μg/ml) 20,25, 30, or 35 days after administration without any other dosageadministration. Preferably, the antibody or antibody fragment has alower EC₅₀ than SYNAGIS®.

[0039] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, said methods comprisingadministering to said mammal a dose of less than 15 mg/kg (preferably 5mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5mg/kg or less) of an antibody or a fragment thereof whichimmunospecifically binds to a RSV antigen and which has an EC₅₀ of lessthan 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM,less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM,less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2nM, in an in vitro microneutralization assay. Preferably, the antibodyor antibody fragment has a lower EC₅₀ than SYNAGIS®.

[0040] The present invention provides methods for achieving atherapeutically or prophylactically effective serum titer in a mammal,said methods comprising administering to said mammal an antibody orfragment thereof which immunospecifically binds to a RSV antigen andwhich has an affinity constant (K_(a)) for a RSV antigen of at least2×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, atleast 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻, at least 10¹¹ M⁻¹, at least 5×10¹¹M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹,or at least 5×10¹⁵ M⁻¹. In particular, the present invention alsoprovides methods for achieving a therapeutically or prophylacticallyeffective serum titer, wherein said effective serum titer is less than30 μg/ml (and is preferably at least 2 μg/ml, more preferably at least 4μg/ml, and most preferably at least 6 μg/ml) after a certain number ofdays (for example, but not limited to, 20, 25, 30 or 35 days) withoutany other dosing within that period, comprising administering to amammal an antibody or fragment thereof that has an affinity constant(K_(a)) for a RSV antigen of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or atleast 5×10¹⁵ M⁻¹. Preferably, the antibody or antibody fragment has ahigher affinity for a RSV F glycoprotein than SYNAGIS®.

[0041] The present invention also provides methods of achieving atherapeutically or prophylactically effective serum titer, wherein saideffective serum titer is less than 30 μg/ml (and is preferably at least2 μg/ml, more preferably at least 4 μg/ml, and most preferably at least6 μg/ml) after a certain number of days (for example, but not limitedto, 20, 25, 30 or 35 days) without any other dosing within that period,comprising administering to a mammal an antibody or fragment thereofwhich immunospecifically binds to a RSV antigen with a higher aviditythan known antibodies such as, e.g., SYNAGIS®.

[0042] The present invention also provides methods of neutralizing RSVusing an antibody or fragment thereof that has an affinity constant(K_(a)) for a RSV antigen of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³M⁻¹, at least 5×10¹³M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or atleast 5×10¹⁵ M⁻¹ to achieve a prophylactically or therapeuticallyeffective serum titer, wherein said effective serum titer is less than30 μg/ml (and is at least 2 μg/ml and more preferably at least 6 μg/ml)20, 25, 30, or 35 days after administration without any other dosageadministration. Preferably, the antibody or antibody fragment has ahigher affinity for the RSV F glycoprotein than SYNAGIS®. The presentinvention also provides methods of neutralizing RSV using an antibody orfragment thereof that has a higher avidity than known antibodies suchas, e.g., SYNAGIS®.

[0043] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, said methods comprisingadministering to said mammal a dose of less than 15 mg/kg (preferably 5mg/kg or less, more preferably 3 mg/kg or less, and most preferably 1.5mg/kg or less) of an antibody or fragment thereof that has an affinityconstant (K_(a)) for a RSV antigen of at least 2×10⁸ M⁻¹, at least2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹ atleast 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³M⁻¹, at least5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻, at least 10¹⁵ M⁻¹, orat least 5×10¹⁵ M⁻¹. Preferably, the antibody or antibody fragment has ahigher affinity for the RSV F glycoprotein than SYNAGIS®. The presentinvention also provides methods for preventing, treating or amelioratingone or more symptoms associated with a RSV infection in a mammal,preferably a human, said methods comprising administering to said mammala first dose of less than 15 mg/kg (preferably 5 mg/kg or less, morepreferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) of anantibody or fragment thereof that has a higher avidity than knownantibodies such as, e.g., SYNAGIS®.

[0044] The present invention encompasses methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, comprising administering tosaid mammal a first dose of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens with higher avidityand/or higher affinity than known antibodies such as, e.g., SYNAGIS®,wherein said effective amount is less than 15 mg/kg (preferably 5 mg/kgor less, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kgor less) of said antibodies or antibody fragments which dose results ina serum titer of less than 30 μg/ml (which is preferably at least 2μg/ml, more preferably at least 4 μg/ml, and most preferably at least 6μg/ml) at least 20 days (preferably at least 25, at least 30, or atleast 35 days) after the administration of the first dose and prior tothe administration of a subsequent dose. In particular, the presentinvention provides methods for preventing, treating, or ameliorating oneor more symptoms associated with a RSV infection in a human subject,comprising administering to said human subject a first dose of less than5 mg/kg (preferably 3 mg/kg or less, and most preferably 1.5 mg/kg) ofan antibody or fragment thereof that immunospecifically binds to a RSVantigen with higher avidity and/or higher affinity than known antibodiessuch as, e.g., SYNAGIS® (e.g., an affinity of at least 2×10⁸ M⁻¹, atleast 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹) so that saidhuman subject has a serum antibody titer of less than 30 μg/ml (which ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) at least 20 days (preferably at least 25,at least 30, or at least 35 days) after the administration of the firstdose of the antibody or antibody fragment and prior to theadministration of a subsequent dose.

[0045] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, said methods comprising administering to saidmammal a first dose of one or more antibodies or fragments thereofcomprising a VH domain having an amino acid sequence of any VH domainlisted in Table 2 to achieve a therapeutically or prophylacticallyeffective serum titer, wherein said effective serum titer is less than30 μg/ml (and is preferably at least 2 μg/ml, more preferably at least 4μg/ml, and most preferably at least 6 μg/ml) after a certain number ofdays (for example, but not limited to, 20, 25, 30 or 35 days) withoutany other dosing within that period. The present invention also providesmethods for preventing, treating or ameliorating one or more symptomsassociated with a RSV infection in a mammal, said methods comprisingadministering to said mammal a first dose of one or more antibodies orfragments thereof comprising one or more VH complementarity determiningregions (CDRs) having the amino acid sequence of one or more VH CDRslisted in Table 2 and/or Table 3 to achieve a therapeutically orprophylactically effective serum titer, wherein said effective serumtiter is less than 30 μg/ml (and is preferably at least 2 μg/ml, morepreferably at least 4 μg/ml, and most preferably at least 6 μg/ml) aftera certain number of days (for example, but not limited to, 20, 25, 30 or35 days) without any other dosing within that period. Preferably, saidantibodies or antibody fragments immunospecifically bind to a RSVantigen.

[0046] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, said methods comprising administering to saidmammal a first dose of one or more antibodies or fragments thereofcomprising a VL domain having the amino acid sequence of any VL domainlisted in Table 2 to achieve a therapeutically or prophylacticallyeffective serum titer, wherein said effective serum titer is less than30 μg/ml (and is preferably at least 2 μg/ml, more preferably at least 4μg/ml, and most preferably at least 6 μg/ml) after a certain number ofdays (for example, but not limited to, 20, 25, 30 or 35 days) withoutany other dosing within that period. The present invention also providesmethods for preventing, treating or ameliorating one or more symptomsassociated with a RSV infection in a mammal, said methods comprisingadministering to said mammal a first dose of one or more antibodies orfragments thereof comprising one or more VL CDRs having the amino acidsequence of one or more VL CDRs listed in Table 2 and/or Table 3 toachieve a therapeutically or prophylactically effective serum titer,wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) after a certain number of days (forexample, but not limited to, 20, 25, 30 or 35 days) without any otherdosing within that period. Preferably, said antibodies or antibodyfragments immunospecifically bind to a RSV antigen.

[0047] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, said methods comprising administering to saidmammal a first dose of one or more antibodies or fragments thereofcomprising a VH domain and a VL domain having the amino acid sequence ofany VH domain and VL domain listed in Table 2 to achieve atherapeutically or prophylactically effective serum titer, wherein saideffective serum titer is less than 30 μg/ml (and is preferably at least2 μg/ml, more preferably at least 4 μg/ml, and most preferably at least6 μg/ml) after a certain number of days (for example, but not limitedto, 20, 25, 30 or 35 days) without any other dosing within that period.The present invention also provides methods for preventing, treating orameliorating one or more symptoms associated with a RSV infection in amammal, said methods comprising administering to said mammal a firstdose of one or more antibodies or fragments thereof comprising one ormore VH CDRs and one or more VL CDRs having the amino acid sequence ofone or more VH CDRs and one or more VL CDRs listed in Table 2 and/or 3to achieve a therapeutically or prophylactically effective serum titer,wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) after a certain number of days (forexample, but not limited to, 20, 25, 30 or 35 days) without any otherdosing within that period. Preferably, said antibodies or antibodyfragments immunospecifically bind to a RSV antigen.

[0048] In a specific embodiment, the present invention provides methodsfor preventing, treating or ameliorating one or more symptoms associatedwith a RSV infection in a mammal, said methods comprising administeringto said mammal a first dose of one or more antibodies or fragmentsthereof comprising a VH domain having an amino acid sequence of SEQ IDNO: 7, 9, 17, 24, 28, 33, 36, 40, 44, 48, 51, 67, or 78 and/or a VLdomain having an amino acid sequence of SEQ ID NO: 8, 11, 13, 21, 26,30, 34, 38, 42, 46, 49, 52, 54, 56, 58, 60, 62, 64, 65, 68, 70, 71, 74or 76 to achieve a therapeutically or prophylactically effective serumtiter, wherein said effective serum titer is less than 30 μg/ml (and ispreferably at least 2 μg/ml, more preferably at least 4 μg/ml, and mostpreferably at least 6 μg/ml) after a certain number of days (forexample, but not limited to, 20, 25, 30 or 35 days) without any otherdosing within that period. In a preferred embodiment, the presentinvention provides methods for preventing, treating or ameliorating oneor more symptoms associated with a RSV infection in a mammal, saidmethods comprising administering to said mammal a first dose of one ormore antibodies or fragments thereof comprising a VH domain having anamino acid sequence of SEQ ID NO: 9, 17, 24, 28, 33, 36, 40, 44, 48, 51,55, 67 or 78 and/or a VL domain having an amino acid sequence of SEQ IDNO: 13, 21, 26, 30, 34, 38, 42, 46, 49, 52, 54, 56, 58, 60, 62, 64, 65,68, 70, 71, 74 or 76 to achieve a therapeutically or prophylacticallyeffective serum titer, wherein said effective serum titer is less than30 μg/ml (and is preferably at least 2 μg/ml, more preferably at least 4μg/ml, and most preferably at least 6 μg/ml) after a certain number ofdays (for example, but not limited to, 20, 25, 30 or 35 days) withoutany other dosing within that period. In another embodiment, the presentinvention provides methods for preventing, treating or ameliorating oneor more symptoms associated with a RSV infection in a mammal, saidmethods comprising administering to said mammal a first dose of one ormore antibodies or fragments thereof comprising a VH CDR3 having anamino acid sequence of SEQ ID NO: 3, 12, 20, 29, or 79 and a VL CDR3having an amino acid sequence of SEQ ID NO: 6, 16 or 61 to atherapeutically or prophylactically effective serum titer, wherein saideffective serum titer is less than 30 μg/ml (and is preferably at least2 μg/ml, more preferably at least 4 μg/ml, and most preferably at least6 82 g/ml) after a certain number of days (for example, but not limitedto, 20, 25, 30 or 35 days) without any other dosing within that period.

[0049] The present invention also provides compositions comprising oneor more antibodies or fragments thereof which immunospecifically bind toone or more RSV antigens and which have increased in vivo half-livescompared to known anti-RSV antibodies as a result of, e.g., one or moremodifications in amino acid residues identified to be involved in theinteraction between the Fc domain of said antibodies or antibodyfragments and the FcRn receptor. In one embodiment, a composition of theinvention comprises HL-SYNAGIS or an antigen-binding fragment thereof.In another embodiment, a composition of the invention comprises one ormore antibodies or fragments thereof which immunospecifically bind toone or more RSV antigens with a higher avidity and/or a higher affinitythan known antibodies such as, e.g., SYNAGIS® (e.g., antibodies orantibody fragments with an affinity of at least 2×10⁸ M⁻¹, at least2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹,at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen)and which comprise an Fc domain with increased affinity for the FcRnreceptor relative to the Fc domain of SYNAGIS®. In accordance with thisembodiment, the increased affinity of the Fc domain of said antibodiesor antibody fragments results in an in vivo half-life of said antibodiesor antibody fragments of at least 25 days, preferably at least 30 days,more preferably at least 30 days, and most preferably at least 40 days.In another embodiment, a composition of the invention comprisesHL-SYNAGIS or an antigen-binding fragment thereof and one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens and which comprise an Fc domain with increasedaffinity for the FcRn receptor relative to the Fc domain of SYNAGIS®.

[0050] The present invention also provides compositions comprising oneor more pegylated antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens. In one embodiment,a composition of the invention comprises pegylated SYNAGIS® or afragment thereof. In another embodiment, a composition of the inventioncomprises one or more pegylated antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens with higher avidityand/or higher affinity than known antibodies such as, e.g., SYNAGIS®. Inyet another embodiment, a composition of the invention comprisespegylated SYNAGIS® or an antigen-binding fragment thereof and one ormore pegylated antibodies or fragments thereof that immunospecificallybind to one or more RSV antigens with higher avidity and/or higheraffinity than known antibodies such as, e.g., SYNAGIS®.

[0051] The present invention also provides compositions comprising oneor more pegylated antibodies or fragments thereof which comprise an Fcdomain with increased affinity for the FcRn receptor relative to the Fcdomain of SYNAGIS®. In one embodiment, a composition of the inventioncomprises a pegylated HL-SYNAGIS or an antigen-binding fragment thereof.In another embodiment, a composition of the invention comprises one ormore pegylated antibodies or fragments thereof which immunospecificallybind to one or more RSV antigens with a higher avidity and/or a higheraffinity than known such as, e.g., SYNAGIS® and which comprise an Fcdomain with increased affinity for the FcRn receptor relative to the Fcdomain of SYNAGIS®

[0052] The present invention encompasses methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, comprising administering tosaid mammal a first dose of a prophylactically or therapeuticallyeffective amount of HL-SYNAGIS or an antigen-binding fragment thereof,wherein said effective amount is approximately 15 mg/kg of saidantibodies or fragments thereof which dose results in a serum titer ofat least 30 μg/ml at least 30 days after the administration of the firstdose and prior to the administration of a subsequent dose. Inparticular, the present invention provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a human subject, comprising administering to said humansubject a first dose of 15 mg/kg of HL-SYNAGIS or an antigen-bindingfragment thereof so that said human subject has a serum antibody titerof at least 30 μg/ml at least 30 days after the administration of thefirst dose of the antibody or antibody fragment and prior to theadministration of a subsequent dose.

[0053] The present invention also encompasses methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, comprising administering tosaid mammal a first dose of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof whichhave increased in vivo half-lives and which immunospecifically bind toone or more RSV antigens with higher avidity and/or higher affinity thanknown antibodies such as, e.g., SYNAGIS® (e.g., antibodies or antibodyfragments with an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen), wherein saideffective amount is less than 15 mg/kg (preferably 5 mg/kg or less, morepreferably 3 mg/kg or less, and most preferably 1.5 mg/kg or less) ofsaid antibodies or fragments thereof which dose results in a serum titerof less than 30 μg/ml (which is preferably at least 2 μg/ml, morepreferably at least 4 μg/ml, and most preferably at least 6 μg/ml) atleast 20 days (preferably at least 25, at least 30, or at least 35 days)after the administration of the first dose and prior to theadministration of a subsequent dose. In particular, the presentinvention provides methods for preventing, treating or ameliorating oneor more symptoms associated with a RSV infection in a human subject,comprising administering to said human subject a first dose of less than5 mg/kg (preferably 1.5 mg/kg or less) of an antibody or a fragmentthereof which has an increased in vivo half-life and whichimmunospecifically binds to a RSV antigen with higher avidity and/orhigher affinity than known antibodies such as, e.g, SYNAGIS® (e.g,antibodies or antibody fragments with an affinity of at least 2×10⁸ M⁻¹,at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, atleast 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSVantigen) so that said human subject has a serum antibody titer of lessthan 30 μg/ml (which is preferably at least 2 μg/ml, more preferably atleast 4 μg/ml, and most preferably at least 6 μg/ml) at least 25 days(preferably at least 30, at least 35, or at least 40 days) after theadministration of the first dose of the antibody or antibody fragmentand prior to the administration of a subsequent dose.

[0054] The present invention provides sustained release formulationscomprising one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens. In one embodiment,a sustained release formulation comprises SYNAGIS® or a fragmentthereof. In another embodiment, a sustained release formulationcomprises one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens with higher avidityand/or higher affinity than known antibodies such as, e.g., SYNAGIS®(e.g., antibodies or antibody fragments with an affinity of at least2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹,at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹for a RSV antigen). In another embodiment, a sustained releaseformulation comprises SYNAGIS® or an antigen-binding fragment thereofand one or more antibodies or fragments thereof that immunospecificallybind to one or more RSV antigens with higher avidity and/or higheraffinity than known antibodies such as, e.g., SYNAGIS® (e.g., antibodiesor antibody fragments with an affinity of at least 2×10⁸ M⁻¹, at least2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹,at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSVantigen). In another embodiment, HL-SYNAGIS or an antigen-bindingfragment thereof is formulated in as sustained release formulation. Inyet another embodiment, antibodies or fragments thereof which havehigher avidity and/or higher affinity for one or more RSV antigens thanknown antibodies such as, e.g., SYNAGIS® (e.g., antibodies or antibodyfragments with an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen) and whichcomprises an Fc domain with increased affinity for the FcRn receptorrelative to the Fc domain of SYNAGIS® are formulated in sustainedrelease formulations.

[0055] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, preferably a human, comprising administering tosaid mammal a first dose of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens in a sustainedrelease formulation, wherein said effective amount is a dose of 15 mg/kgor less of said antibodies or fragments thereof, which dose, preferablyresults in a serum titer of at least 2 μg/ml (preferably at least 5μg/ml, at least 10 μg/ml, at least 20 μg/ml, at least 30 μg/ml, or atleast 40 μg/ml) for at least 20 days (preferably at least 25, 30, 35 or40 days) after the administration of the first dose and prior to theadministration of a subsequent dose.

[0056] In one embodiment, a mammal, preferably a human, is administereda first dose of a prophylactically or therapeutically effective amountof SYNAGIS® or an antigen-binding fragment thereof in a sustainedrelease formulation, wherein said effective amount is a dose ofapproximately 15 mg/kg of SYNAGIS® or an antigen-binding fragmentthereof which dose results in a serum titer of at least 20 μg/ml(preferably at least 30 μg/ml, more preferably at least 40 μg/ml, andmost preferably at least 50 μg/ml) for at least 30 days (preferably atleast 35 days, more preferably at least 40 days, and most preferably atleast 45 days) after the administration of the first dose and prior tothe administration of a subsequent dose. In a preferred embodiment, amammal, preferably a human, is administered a first dose of aprophylactically or therapeutically effective amount of SYNAGIS® or anantigen-binding fragment thereof in a sustained release formulation,wherein said effective amount is a dose of 15 mg/kg or less of SYNAGIS®or an antigen-binding fragment thereof which dose results in a serumtiter of 20 μg/ml (preferably at least 30 μg/ml, more preferably atleast 40 μg/ml, and most preferably at least 50 μg/ml) at least 30 days(preferably at least 35 days, more preferably at least 40 days, and mostpreferably at least 45 days) after the administration of the first doseand prior to the administration of a subsequent dose.

[0057] In another embodiment, a mammal, preferably a human, isadministered a first dose of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens with higher avidityand/or higher affinity than known antibodies such as, e.g., SYNAGIS®(e.g., antibodies or antibody fragments with an affinity of at least2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹,at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹for a RSV antigen) in a sustained release formulation, wherein saideffective amount is a dose of less than 15 mg/kg (preferably 5 mg/kg orless, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg orless) of said antibodies or antibody fragments which dose results in aserum titer of less than 30 μg/ml (which is preferably at least 2 μg/ml,more preferably at least 4 μg/ml, and most preferably at least 6 μg/ml)for at least 20 days (preferably at least 25, at least 30, at least 35,or at least 40 days) after the administration of the first dose andprior to the administration of a subsequent dose. In a preferredembodiment, a mammal, preferably a human, is administered a first doseof a prophylactically or therapeutically effective amount of one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens with higher avidity and/or higher affinity than knownantibodies such as, e.g, SYNAGIS® (e.g., antibodies or antibodyfragments with an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10 M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen) in a sustainedrelease formulation, wherein said effective amount is a dose of lessthan 15 mg/kg of said antibodies or antibody fragments which doseresults in a serum titer of 10 μg/ml for at least 20 days (preferably atleast 25, at least 30, at least 35 or at least 40 days) after theadministration of the first dose and prior to the administration of asubsequent dose. In accordance with this embodiment, theprophylactically or therapeutically effective amount of the dose of theantibodies or antibody fragments is approximately 0.5 mg/kg, preferably1 mg/kg, 1.5 mg/kg, 3 mg/kg, 5 mg/kg, 7.5 mg/kg, 10 mg/kg, 12 mg/kg, or14 mg/kg. In another preferred embodiment, a mammal, preferably a human,is administered a first dose of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens with higher avidityand/or higher affinity than known antibodies such as, e.g., SYNAGIS®(e.g., antibodies or antibody fragments with an affinity of at least2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹,at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹for a RSV antigen) in a sustained release formulation, wherein saideffective amount is a dose of 1.5 mg/kg of said antibodies or antibodyfragments which dose results in a serum titer of 10 μg/ml for at least20 days (preferably at least 25, at least 30, at least 35, or at least40 days) after the administration of the first dose and prior to theadministration of a subsequent dose.

[0058] Additionally, the present invention provides sustained releasecompositions comprising one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens, whichsustained release compositions maintain a certain serum titer in asubject for a certain period of time without exceeding a particularserum titer. In one embodiment, a sustained release formulationcomprising SYNAGIS® or an antigen-binding fragment thereof maintains aserum titer in a mammal, preferably a human, of approximately 25 μg/ml(preferably 30 μg/ml, more preferably 40 μg/ml, and most preferably 50μg/ml) without exceeding a serum titer of approximately 100 μg/ml(preferably 75 μg/ml) for at least 20 days (preferably at least 25, 30,35, or 40 days). In another embodiment, a sustained release formulationcomprising one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens with a higheravidity and/or a higher affinity than previously known antibodies suchas, e.g., SYNAGIS®, maintains a serum titer in a mammal, preferably ahuman, of approximately 2 μg/ml (preferably 6 μg/ml, 10 μg/ml, 20 μg/ml,or 30 μg/ml) without exceeding a serum titer of approximately 40 μg/ml(preferably 75 μg/ml) for at least 20 days (preferably at least 25, 30,35, or 40 days).

[0059] The present invention encompasses methods of preventing, treatingor ameliorating one or more symptoms of RSV infection in a mammal,preferably a human, by administering sustained release formulations ofone or more antibodies or fragments thereof which immunospecificallybind to one or more RSV antigens and which have increased in vivohalf-lives. In one embodiment, a sustained release formulationcomprising HL-SYNAGIS or an antigen-binding fragment thereof isadministered to a mammal, preferably a human, to prevent, treat, orameliorate one or more symptoms associated with a RSV infection. Inanother embodiment, a sustained release formulation comprising one ormore antibodies or fragments thereof which have higher avidity and/orhigher affinity for one or more RSV antigens than known antibodies suchas, e.g., SYNAGIS® (e.g., antibodies or antibody fragments with anaffinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹,or at least 5×10¹² M⁻¹ for a RSV antigen) and which comprises an Fcdomain with increased affinity for the FcRn receptor relative to the Fcdomain of SYNAGIS® are administered to a mammal, preferably a human, toprevent, treat, or ameliorate one or more symptoms associated with a RSVinfection.

[0060] The present invention also provides pulmonary delivery systemsfor administering one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens. In particular, thepresent invention provides compositions for pulmonary delivery, saidcompositions comprising one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens. SYNAGIS® oran antigen-binding fragment thereof can be incorporated intocompositions for pulmonary delivery. HL-SYNAGIS or an antigen-bindingfragment thereof can be incorporated into compositions for pulmonarydelivery. One or more antibodies or fragments thereof that bind to oneor more RSV antigens with higher affinity and/or higher avidity thanknown antibodies such as, e.g., SYNAGIS® (e.g., antibodies or antibodyfragments with an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen) can beincorporated into compositions for pulmonary delivery. Further, one ormore antibodies or fragments thereof which bind to one or more RSVantigens with higher affinity and/or higher avidity than knownantibodies such as, e.g., SYNAGIS(& (e.g., antibodies or antibodyfragments with an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for a RSV antigen) and whichcomprise an Fc domain with increased affinity for the FcRn receptorrelative to the Fc domain of SYNAGIS® can be incorporated intocompositions for pulmonary delivery.

[0061] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms associated with a RSVinfection, said methods comprising administering to a mammal, preferablya human, a composition for pulmonary delivery comprising one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens. In particular, the present invention provides methodsfor preventing, treating or ameliorating one or more symptoms associatedwith a RSV infection, said methods comprising administering to a mammal,preferably a human, a composition for pulmonary delivery comprisingSYNAGIS® or fragments thereof. The present invention also providesmethods for preventing, treating or ameliorating one or more symptomsassociated with a RSV infection, said methods comprising administeringto a mammal, preferably a human, a composition for pulmonary deliverycomprising one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens with higher affinityand/or higher avidity than known antibodies such as, e.g., SYNAGIS®(e.g., antibodies or antibody fragments having an affinity of at least2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹,at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹for one or more RSV antigens).

[0062] In one embodiment, a first dose of a prophylactically ortherapeutically effective amount of a composition comprising SYNAGIS® oran antigen-binding fragment thereof is administered to the lungs of amammal, preferably a human, and results in an antibody concentration ofat least 20 ng per mg of lung protein (preferably at least 40 ng/mg, atleast 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, at least 75 ng/mg,at least 100 ng/mg, or at least 150 ng/mg) at least 20 days (preferablyat least 25, 30, 35 or 40 days) after the administration of the firstdose and prior to the administration of a subsequent dose. Preferably,the prophylactically or therapeutically effective amount is a dose ofapproximately 0.01 mg/kg, (preferably at least 0.1 mg/kg, at least 1mg/kg, at least 2 mg/kg, at least 4 mg/kg, at least 5 mg/kg or at least10 mg/kg) of SYNAGIS® or an antigen-binding fragment thereof.

[0063] In another embodiment, a first dose of a prophylactically ortherapeutically effective amount of a composition comprising one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens with higher affinity and/or higher avidity than knownantibodies such as, e.g., SYNAGIS® (e.g., antibodies or antibodyfragments having an affinity of at least 2×10⁸ M⁻¹, at least 2.5×10⁸M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹,at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ for one or more RSV antigens)is administered to the lungs of a mammal, preferably a human and resultsin an antibody concentration of 20 ng per mg of lung protein (preferablyat least 40 ng/mg, at least 60 ng/mg, at least 80 ng/mg, at least 50ng/mg, at least 75 ng/mg, at least 100 ng/mg, or at least 150 ng/mg) ,at least 200 ng/mg, at least 250 ng/mg, at least 500 ng/mg, at least 750ng/mg, at least 1 μg/mg, at least 2 μg/mg, at least 5 μg/mg, at least 10μg/mg, at least 15 μg/mg, or at least 25 μg/mg) at least 20 days(preferably at least 25, 30, 35 or 40 days) at least 20 days (preferablyat least 25, at least 30, at least 35 or at least 40 days after theadministration of the first dose and prior to the administration of asubsequent dose. Preferably, the prophylactically effective amount is adose of approximately 0.001 mg/kg, (preferably at least 0.005 mg/kg, atleast 0.01 mg/kg, at least 0.05 mg/kg, at least 0.1 mg/kg, at least 1mg/kg, at least 2 mg/kg, at least 4 mg/kg, at least 5 mg/kg or at least10 mg/kg) of said antibodies or antibody fragments.

[0064] The present invention further provides detectable or diagnosticcompositions comprising using antibodies or fragments thereof thatimmunospecifically bind to a RSV antigen, and methods for detecting ordiagnosing a RSV infection utilizing said compositions.

[0065] 3.1. DEFINITIONS

[0066] The term “analog” as used herein refers to a polypeptide thatpossesses a similar or identical function as a RSV polypeptide, afragment of a RSV polypeptide, an antibody, or antibody fragment butdoes not necessarily comprise a similar or identical amino acid sequenceof a RSV polypeptide, a fragment of a RSV polypeptide, an antibody, orantibody fragment, or possess a similar or identical structure of a RSVpolypeptide, a fragment of a RSV polypeptide, an antibody, or antibodyfragment. A polypeptide that has a similar amino acid sequence refers toa polypeptide that satisfies at least one of the following: (a) apolypeptide having an amino acid sequence that is at least 30%, at least35%, at least 40%, at least 45%, at least 50%, at least 55%, at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95% or at least 99% identical to the aminoacid sequence of a RSV polypeptide, a fragment of a RSV polypeptide, anantibody, or antibody fragment described herein; (b) a polypeptideencoded by a nucleotide sequence that hybridizes under stringentconditions to a nucleotide sequence encoding a RSV polypeptide, afragment of a RSV polypeptide, an antibody, or antibody fragmentdescribed herein of at least 5 amino acid residues, at least 10 aminoacid residues, at least 15 amino acid residues, at least 20 amino acidresidues, at least 25 amino acid residues, at least 40 amino acidresidues, at least 50 amino acid residues, at least 60 amino residues,at least 70 amino acid residues, at least 80 amino acid residues, atleast 90 amino acid residues, at least 100 amino acid residues, at least125 amino acid residues, or at least 150 amino acid residues; and (c) apolypeptide encoded by a nucleotide sequence that is at least 30%, atleast 35%, at least 40%, at least 45%, at least 50%, at least 55%, atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95% or at least 99% identical to thenucleotide sequence encoding a RSV polypeptide, a fragment of a RSVpolypeptide, an antibody, or antibody fragment described herein. Apolypeptide with similar structure to a RSV polypeptide, a fragment of aRSV polypeptide, an antibody, or antibody fragment described hereinrefers to a polypeptide that has a similar secondary, tertiary orquaternary structure of a RSV polypeptide, a fragment of a RSV, anantibody, or antibody fragment described herein. The structure of apolypeptide can determined by methods known to those skilled in the art,including but not limited to, X-ray crystallography, nuclear magneticresonance, and crystallographic electron microscopy.

[0067] The term “derivative” as used herein refers to a polypeptide thatcomprises an amino acid sequence of a RSV polypeptide, a fragment of aRSV polypeptide, an antibody that immunospecifically binds to a RSVpolypeptide, or an antibody fragment that immunospecifically binds to aRSV polypeptide which has been altered by the introduction of amino acidresidue substitutions, deletions or additions. The term “derivative” asused herein also refers to a RSV polypeptide, a fragment of a RSVpolypeptide, an antibody that immunospecifically binds to a RSVpolypeptide, or an antibody fragment that immunospecifically binds to aRSV polypeptide which has been modified, i.e, by the covalent attachmentof any type of molecule to the polypeptide. For example, but not by wayof limitation, a RSV polypeptide, a fragment of a RSV polypeptide, anantibody, or antibody fragment may be modified, e.g., by glycosylation,acetylation, pegylation, phosphorylation, amidation, derivatization byknown protecting/blocking groups, proteolytic cleavage, linkage to acellular ligand or other protein, etc. A derivative of a RSVpolypeptide, a fragment of a RSV polypeptide, an antibody, or antibodyfragment may be modified by chemical modifications using techniquesknown to those of skill in the art, including, but not limited tospecific chemical cleavage, acetylation, formylation, metabolicsynthesis of tunicamycin, etc. Further, a derivative of a RSVpolypeptide, a fragment of a RSV polypeptide, an antibody, or antibodyfragment may contain one or more non-classical amino acids. Apolypeptide derivative possesses a similar or identical function as aRSV polypeptide, a fragment of a RSV polypeptide, an antibody, orantibody fragment described herein.

[0068] The term “effective neutralizing titer” as used herein refers tothe amount of antibody which corresponds to the amount present in theserum of animals (human or cotton rat) that has been shown to be eitherclinically efficacious (in humans) or to reduce virus by 99% in, forexample, cotton rats. The 99% reduction is defined by a specificchallenge of, e.g., 10³ pfu, 10⁴ pfu, 10⁵ pfu, 10⁶ pfu, 10⁷ pfu, 10⁸pfu, or 10⁹ pfu) of RSV.

[0069] The term “epitopes” as used herein refers to portions of a RSVpolypeptide having antigenic or immunogenic activity in an animal,preferably a mammal, and most preferably in a human. An epitope havingimmunogenic activity is a portion of a RSV polypeptide that elicits anantibody response in an animal. An eptiope having antigenic activity isa portion of a RSV polypeptide to which an antibody immunospecificallybinds as determined by any method well known in the art, for example, bythe immunoassays described herein. Antigenic epitopes need notnecessarily be immunogenic.

[0070] The term “fragment” as used herein refers to a peptide orpolypeptide comprising an amino acid sequence of at least 5 contiguousamino acid residues, at least 10 contiguous amino acid residues, atleast 15 contiguous amino acid residues, at least 20 contiguous aminoacid residues, at least 25 contiguous amino acid residues, at least 40contiguous amino acid residues, at least 50 contiguous amino acidresidues, at least 60 contiguous amino residues, at least 70 contiguousamino acid residues, at least contiguous 80 amino acid residues, atleast contiguous 90 amino acid residues, at least contiguous 100 aminoacid residues, at least contiguous 125 amino acid residues, at least 150contiguous amino acid residues, at least contiguous 175 amino acidresidues, at least contiguous 200 amino acid residues, or at leastcontiguous 250 amino acid residues of the amino acid sequence of a RSVpolypeptide or an antibody that immunospecifically binds to a RSVpolypeptide.

[0071] The term “human infant” as used herein refers to a human lessthan 24 months, preferably less than 16 months, less than 12 months,less than 6 months, less than 3 months, less than 2 months, or less than1 month of age.

[0072] The term “human infant born prematurely” as used herein refers toa human born at less than 40 weeks gestational age, preferably less than35 weeks gestational age, who is less than 6 months old, preferably lessthan 3 months old, more preferably less than 2 months old, and mostpreferably less than 1 month old.

[0073] An “isolated” or “purified” antibody or fragment thereof issubstantially free of cellular material or other contaminating proteinsfrom the cell or tissue source from which the protein is derived, orsubstantially free of chemical precursors or other chemicals whenchemically synthesized. The language “substantially free of cellularmaterial” includes preparations of an antibody or antibody fragment inwhich the antibody or antibody fragment is separated from cellularcomponents of the cells from which it is isolated or recombinantlyproduced. Thus, an antibody or antibody fragment that is substantiallyfree of cellular material includes preparations of antibody or antibodyfragment having less than about 30%, 20%, 10%, or 5% (by dry weight) ofheterologous protein (also referred to herein as a “contaminatingprotein”). When the antibody or antibody fragment is recombinantlyproduced, it is also preferably substantially free of culture medium,i.e., culture medium represents less than about 20%, 10%, or 5% of thevolume of the protein preparation. When the antibody or antibodyfragment is produced by chemical synthesis, it is preferablysubstantially free of chemical precursors or other chemicals, i.e., itis separated from chemical precursors or other chemicals which areinvolved in the synthesis of the protein. Accordingly such preparationsof the antibody or antibody fragment have less than about 30%, 20%, 10%,5% (by dry weight) of chemical precursors or compounds other than theantibody or antibody fragment of interest. In a preferred embodiment,antibodies of the invention or fragments thereof are isolated orpurified.

[0074] An “isolated” nucleic acid molecule is one which is separatedfrom other nucleic acid molecules which are present in the naturalsource of the nucleic acid molecule. Moreover, an “isolated” nucleicacid molecule, such as a cDNA molecule, can be substantially free ofother cellular material, or culture medium when produced by recombinanttechniques, or substantially free of chemical precursors or otherchemicals when chemically synthesized. In a preferred embodiment,nucleic acid molecules encoding antibodies of the invention or fragmentsthereof are isolated or purified.

[0075] The term “fusion protein” as used herein refers to a polypeptidethat comprises an amino acid sequence of an antibody or fragment thereofand an amino acid sequence of a heterologous polypeptide (e.g., anon-anti-RSV antigen antibody).

[0076] The term “high potency” as used herein refers to antibodies orfragments thereof that exhibit high potency as determined in variousassays for biological activity (e.g., neutralization of RSV) such asthose described herein. For example, high potency antibodies of thepresent invention or fragments thereof have an EC₅₀ value less than 0.01nM, less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM asmeasured by a microneutralization assay described herein. Further, highpotency antibodies of the present invention or fragments thereof resultin at least a 75%, preferably at least a 95% and more preferably a 99%lower RSV titer in a cotton rat 5 days after challenge with 10⁵ pfurelative to a cotton rat not administered said antibodies or antibodyfragments. In certain embodiments of the invention, high potencyantibodies of the present invention or fragments thereof exhibit a highaffinity and/or high avidity for one or more RSV antigens (e.g.,antibodies or antibody fragments having an affinity of at least 2×10⁸M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, atleast 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, or at least 5×10¹² M⁻¹ forone or more RSV antigens).

[0077] The term “host” as used herein refers to a mammal, preferably ahuman.

[0078] The term “host cell” as used herein refers to the particularsubject cell transfected with a nucleic acid molecule and the progeny orpotential progeny of such a cell. Progeny of such a cell may not beidentical to the parent cell transfected with the nucleic acid moleculedue to mutations or environmental influences that may occur insucceeding generations or integration of the nucleic acid molecule intothe host cell genome.

[0079] In certain embodiments of the invention, a “prophylacticallyeffective serum titer” is the serum titer in a mammal, preferably ahuman, that reduces the incidence of a RSV infection in said mammal.Preferably, the prophylactically effective serum titer reduces theincidence of RSV infections in humans with the greatest probability ofcomplications resulting from RSV infection (e.g., a human with cysticfibrosis, bronchopulmonary dysplasia, congenital heart disease,congenital immunodeficiency or acquired immunodeficiency, a human whohas had a bone marrow transplant, a human infant, or an elderly human).In certain other embodiments of the invention, a “prophylacticallyeffective serum titer” is the serum titer in a cotton rat that resultsin a RSV titer 5 days after challenge with 10⁵ pfu that is 99% lowerthan the RSV titer 5 days after challenge with 10⁵ pfu of RSV in acotton rat not administered an antibody or antibody fragment thatimmunospecifically binds to a RSV antigen.

[0080] In certain embodiments of the invention, a “therapeuticallyeffective serum titer” is the serum titer in a mammal, preferably ahuman, that reduces the severity, the duration and/or the symptomsassociated with a RSV infection in said mammal. Preferably, thetherapeutically effective serum titer reduces the severity, the durationand/or the number symptoms associated with RSV infections in humans withthe greatest probability of complications resulting from a RSV infection(e.g., a human with cystic fibrosis, bronchopulmonary dysplasia,congenital heart disease, congenital immunodeficiency or acquiredimmunodeficiency, a human who has had a bone marrow transplant, a humaninfant, or an elderly human). In certain other embodiments of theinvention, a “therapeutically effective serum titer” is the serum titerin a cotton rat that results in a RSV titer 5 days after challenge with10⁵ pfu that is 99% lower than the RSV titer 5 days after challenge with10⁵ pfu of RSV in a cotton rat not administered an antibody or antibodyfragment that immunospecifically binds to a RSV antigen.

[0081] As used herein, “HL-SYNAGIS” is SYNAGIS® with one or moremodifications in amino acid residues identified to be involved in theinteraction between the Fc domain of SYNAGIS® and the FcRn receptorwhich results in an increase in the in vivo half-life of SYNAGIS® togreater than 21 days. An antigen-binding fragment of HL-SYNAGIS is afragment of SYNAGIS® which immunospecifically binds to RSV Fglycoprotein and has one or more modifications in amino acid residuesidentified to be involved in the interaction between the Fc domain ofSYNAGIS® and the FcRn receptor, wherein said modifications result in anincrease in the in vivo half-life of the antigen-binding fragment. Inaccordance with the invention, HL-SYNAGIS or an antigen-binding fragmentthereof has an in vivo half-life of at least 25 days, preferably atleast 30 days, more preferably at least 35 days, and most preferably atleast 40 days.

[0082] The term “RSV antigen” refers to a RSV polypeptide or fragmentthereof to which an antibody or antibody fragment immunospecificallybinds. An RSV antigen also refers to an analog or derivative of a RSVpolypeptide or fragment thereof to which an antibody or antibodyfragment immunospecifically binds.

[0083] The term “serum titer” as used herein refers to an average serumtiter in a population of least 10, preferably at least 20, and mostpreferably at least 40 subjects.

[0084] The term “antibodies or fragments that immunospecifically bind toa RSV antigen” as used herein refers to antibodies or fragments thereofthat specifically bind to a RSV polypeptide or a fragment of a RSVpolypeptide and do not non-specifically bind to other polypeptides.Antibodies or fragments that immunospecifically bind to a RSVpolypeptide or fragment thereof may have cross-reactivity with otherantigens. Preferably, antibodies or fragments that immunospecificallybind to a RSV polypeptide or fragment thereof do not cross-react withother antigens. Antibodies or fragments that immunospecifically bind toa RSV polypeptide can be identified, for example, by immunoassays orother techniques known to those of skill in the art.

[0085] To determine the percent identity of two amino acid sequences orof two nucleic acid sequences, the sequences are aligned for optimalcomparison purposes (e.g., gaps can be introduced in the sequence of afirst amino acid or nucleic acid sequence for optimal alignment with asecond amino acid or nucleic acid sequence). The amino acid residues ornucleotides at corresponding amino acid positions or nucleotidepositions are then compared. When a position in the first sequence isoccupied by the same amino acid residue or nucleotide as thecorresponding position in the second sequence, then the molecules areidentical at that position. The percent identity between the twosequences is a function of the number of identical positions shared bythe sequences (i.e., % identity number of identical overlappingpositions/total number of positions ×100%). In one embodiment, the twosequences are the same length.

[0086] The determination of percent identity between two sequences canalso be accomplished using a mathematical algorithm. A preferred,non-limiting example of a mathematical algorithm utilized for thecomparison of two sequences is the algorithm of Karlin and Altschul,1990, Proc. Natl. Acad. Sci. U.S.A. 87:2264-2268, modified as in Karlinand Altschul, 1993, Proc. Natl. Acad. Sci. U.S.A. 90:5873-5877. Such analgorithm is incorporated into the NBLAST and XBLAST programs ofAltschul et al., 1990, J. Mol. Biol. 215:403. BLAST nucleotide searchescan be performed with the NBLAST nucleotide program parameters set,e.g., for score=100, wordlength=12 to obtain nucleotide sequenceshomologous to a nucleic acid molecules of the present invention. BLASTprotein searches can be performed with the XBLAST program parametersset, e.g., to score-50, wordlength=3 to obtain amino acid sequenceshomologous to a protein molecule of the present invention. To obtaingapped alignments for comparison purposes, Gapped BLAST can be utilizedas described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402.Alternatively, PSI-BLAST can be used to perform an iterated search whichdetects distant relationships between molecules (Id.). When utilizingBLAST, Gapped BLAST, and PSI-Blast programs, the default parameters ofthe respective programs (e.g., of XBLAST and NBLAST) can be used (see,e.g., http://www.ncbi.nlm.nih.gov). Another preferred, non-limitingexample of a mathematical algorithm utilized for the comparison ofsequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17.Such an algorithm is incorporated in the ALIGN program (version 2.0)which is part of the GCG sequence alignment software package. Whenutilizing the ALIGN program for comparing amino acid sequences, a PAM120weight residue table, a gap length penalty of 12, and a gap penalty of 4can be used.

[0087] The percent identity between two sequences can be determinedusing techniques similar to those described above, with or withoutallowing gaps. In calculating percent identity, typically only exactmatches are counted.

4. DESCRIPTION OF THE FIGURES

[0088] FIGS. 1A-1B show the amino acid sequences of the (A) light chainvariable region and (B) heavy chain variable region of a high affinitymonoclonal antibody that binds to a RSV antigen the potency of which canbe increased by methods described herein or in Applicants' copendingapplication Ser. Nos. 60/168,426 and 60/186,252. For reference purposes,this is the amino acid sequence of the SYNAGIS® antibody disclosed inJohnson et al, 1997, J. Infect. Dis. 176:1215-1224 and U.S. Pat. No.5,824,307. Here, the CDR regions are underlined while non-underlinedresidues form the framework regions of the variable regions of eachantibody. In this antibody, the CDRs are derived from a mouse antibodywhile the framework regions are derived from a human antibody. Theconstant regions (not shown) are also derived from a human antibody.

[0089] FIGS. 2A-2B show the (A) light chain variable region and (B)heavy light chain variable region for an antibody sequence. CDR regionsare underlined. This sequence differs from the sequence disclosed inFIGS. 1A-1B in the first 4 residues of VH CDR1 of the light chain,residue 103 of the light chain and residue 112 of the heavy chain.

5. DETAILED DESCRIPTION OF THE INVENTION

[0090] The present invention provides methods of preventing,neutralizing, treating and ameliorating one or more symptoms associatedwith a RSV infection in a subject comprising administering to saidsubject one or more antibodies which immunospecifically bind to one ormore RSV antigens with high affinity and/or high avidity and/or have alonger serum half-life. The high affinity and/or high avidity of theantibodies of the invention enable the use of lower doses of saidantibodies than previously thought to be effective for the prevention,neutralization, treatment and the amelioration of symptoms associatedwith RSV infection. The use of lower doses of antibodies whichimmunospecifically bind to one or more RSV antigens reduces thelikelihood of adverse effects, as well as providing a more effectiveprophylaxis. Further, the high affinity and/or high avidity of theantibodies of the invention enable less frequent administration of saidantibodies than previously thought to be necessary for the prevention,neutralization, treatment and the amelioration of symptoms associatedwith RSV infection.

[0091] The present invention also provides methods of preventing,neutralizing, treating and ameliorating one or more symptoms associatedwith a RSV infection in a subject comprising administering to saidsubject one or more antibodies which immunospecifically bind to one ormore RSV antigens, said antibodies having a longer half-life than otherpreviously known antibodies.

[0092] The present invention also provides improved methods ofadministering one or more antibodies which immunospecifically bind toone or more RSV antigens to a subject, said methods enable lower dosesof said antibodies to be administered to the subject while achievingserum titers effective for the prevention, neutralization, treatment andamelioration of one or more symptoms associated with RSV infection. Thepresent invention encompasses methods of delivering one or moreantibodies which immunospecifically bind to one or more RSV antigensdirectly to the site of RSV infection. In particular, the inventionencompasses pulmonary delivery of one or more antibodies whichimmunospecifically bind to one or more RSV antigens. The improvedmethods of delivering of one or more antibodies which immunospecificallybind to one or more RSV antigens reduces the dosage and frequency ofadministration of said antibodies to a subject.

[0093] The present invention is based, in part, upon achieving orinducing a serum titer of 1 μg/ml or less, preferably 2 μg/ml or less, 5μg/ml or less, 6 μg/ml or less, 10 μg/ml or less, 15 μg/ml or less, 20μg/ml or less, or 25 μg/ml or less of an antibody or fragment thereofthat immunospecifically binds to a respiratory syncytial virus (RSV)antigen in a mammal with higher affinity and/or higher avidity thanpreviously known antibodies, while reducing or avoiding adverse affects.Preferably a serum titer or serum titer of 1 μg/ml or less, preferably 2μg/ml or less, 5 μg/ml or less, 6 μg/ml or less, 10 μg/ml or less, 15μg/ml or less, 20 μg/ml or less, or 25 μg/ml or less is achievedapproximately 20 days (preferably 25, 30, 35 or 40 days) afteradministration of a first dose of antibodies or fragments thereof whichimmunospecifically bind to a RSV antigen and without administration ofany other doses of said antibodies or fragments thereof.

[0094] The present invention provides methods of achieving or inducing aserum titer of at least 30 μg/ml, at least 40 μg/ml, at least 50 μg/ml,at least 75 μg/ml, at least 100 μg/ml, at least 125 μ/ml, at least 150μg/ml, at least 175 μg/ml, at least 200 μg/ml, at least 225 μg/ml, atleast 250 μg/ml, at least 275 μg/ml, at least 300 μg/ml, at least 325μg/ml, at least 350 μg/ml, at least 375 μg/ml, or at least 400 μg/ml ofan antibody or fragment thereof that immunospecifically binds to arespiratory syncytial virus (RSV) antigen in a mammal, while reducing oravoiding adverse affects. Preferably a serum titer or serum titer of atleast 30 μg/ml, preferably at least 40 μg/ml, at least 50 μg/ml, atleast 75 μg/ml, at least 100 μg/ml, at least 125 μg/ml, at least 150μg/ml, at least 175 μg/ml, at least 200 μg/ml, at least 225 μg/ml, atleast 250 μg/ml, at least 275 μg/ml, at least 300 μg/ml, at least 325μg/ml, at least 350 μg/ml, at least 375 μg/ml, or at least 400 μg/ml isachieved approximately 30 days after administration of a first dose ofantibodies or fragments thereof which immunospecifically bind to a RSVantigen and without administration of any other doses of said antibodiesor fragments thereof.

[0095] In a specific embodiment, a serum titer in a non-primate mammalof at least 40 μg/ml, preferably at least 80 μg/ml, at least 100 μg/ml,at least 120 μg/ml, at least 150 μg/ml, at least 200 μg/ml, at least 250μg/ml, or at least 300 μg/ml, of one or more antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens isachieved at least 1 day after administering a dose of less than 2.5mg/kg, preferably less than 1 mg/kg, or less than 0.5 mg/kg of theantibodies or antibody fragments to the non-primate mammal. In anotherembodiment, a serum titer in a non-primate mammal of at least 150 μg/ml,preferably at least 200 μg/ml, at least 250 μg/ml, at least 300 μg/ml,at least 350 μg/ml, or at least 400 μg/ml of one or more antibodies orfragments thereof that immunospecifically bind to one or more RSVantigens is achieved at least 1 day after administering a dose ofapproximately 5 mg/kg of the antibodies or antibody fragments to thenon-primate mammal.

[0096] In another embodiment, a serum titer in a primate of at least 40μg/ml, preferably at least 80 μg/ml, at least 100 μg/ml, at least 120μg/ml, at least 150 μg/ml, at least 200 μg/ml, at least 250 μg/ml, or atleast 300 μg/ml of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens is achieved at least30 days after administering a first dose of less than 5 mg/kg,preferably less than 3 mg/kg, less than 1 mg/kg, or less than 0.5 mg/kgof the antibodies or fragments thereof to the primate. In yet anotherembodiment, a serum titer in a primate of at least 200 μg/ml, at least250 μg/ml, at least 300 μg/ml, at least 350 μg/ml, or at least 400 μg/mlof one or more antibodies or fragments thereof that immunospecificallybind to one or more RSV antigens is achieved at least 30 days afteradministering a first dose of approximately 15 mg/kg of the antibodiesor fragments thereof to the primate. In accordance with theseembodiments, the primate is preferably a human.

[0097] The present invention provides methods for preventing, treating,or ameliorating one or more symptoms associated with a RSV infection ina mammal, preferably a human, said methods comprising administering afirst dose to said mammal of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens, wherein saideffective amount is less than 15 mg/kg of said antibodies or fragmentsthereof and which results in a serum titer of greater than 40 μg/ml 30days after the first administration and prior to any subsequentadministration. In one embodiment, RSV infection in a human subject isprevented or treated, or one or more symptoms associated with RSVinfection is ameliorated by administering a first dose of less than 10mg/kg, preferably less than 5 mg/kg, less than 3 mg/kg, or less than 1mg/kg of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens so that a serumantibody titer of at least 40 μg/ml, preferably at least 80 μg/ml, or atleast 120 μg/ml, at least 150 μg/ml, at least 200 μg/ml, at least 250μg/ml, or at least 300 μg/ml is achieved 30 days after theadministration of the first dose of the antibodies or antibody fragmentsand prior to the administration of a subsequent dose. In anotherembodiment, RSV infection in a human subject is prevented or treated, orone or more symptoms associated with a RSV infection is ameliorated byadministering a first dose of approximately 15 mg/kg of one or moreantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens so that a serum antibody titer of at least 75 μg/ml,preferably at least 100 μg/ml, at least 200 μg/ml, at least 250 μ/ml, atleast 300 μg/ml, at least 350 μg/ml, or at least 400 μg/ml is achieved30 days after the administration of the first dose of the antibodies orantibody fragments and prior to the administration of a subsequent dose.In yet another embodiment, RSV infection in a human subject is preventedor treated, or one or more symptoms associated with a RSV infection isameliorated by administering a first dose of one or more antibodies orfragments thereof that immunospecifically bind to one or more RSVantigens such that a prophylactically or therapeutically effective serumtiter of less than 10 μg/ml, preferably less than 5 μg/ml, less than 3μg/ml, less than 1 μg/ml, or less than 0.5 μg/ml is achieved no morethan 30 days after administering the antibodies or antibody fragments.In accordance with this embodiment, the first dose of one or moreantibodies or fragments thereof is less than 10 mg/kg, preferably lessthan 5 mg/kg, less than 1 mg/kg, or less than 0.5 mg/kg.

[0098] The present invention provides antibodies or fragments thereofwhich immunospecifically bind to a RSV antigen with an affinity constantof at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, atleast 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹,at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹.Preferably, the antibodies or antibody fragments have a higher affinityfor a RSV antigen than SYNAGIS® does for the RSV F glycoprotein. Thepresent invention also provides pharmaceutical compositions comprisingone or more antibodies which immunospecifically bind to a RSV antigenwith an affinity constant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹,at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, atleast 10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or atleast 5×10¹⁵ M⁻¹.

[0099] The present invention also provides antibodies or fragmentsthereof which immunospecifically bind to a RSV antigen with a higheravidity than any previously known antibodies or fragments thereof.Preferably, the antibodies or antibody fragments have higher avidity fora RSV antigen than SYNAGIS® has for the RSV F glycoprotein. The presentinvention also provides antibodies or fragments thereof thatimmunospecifically bind to a RSV antigen which have a higher affinityfor a RSV antigen than any previously known antibodies or fragmentsthereof. The present invention also provides pharmaceutical compositionscomprising one or more antibodies or fragments thereof whichimmunospecifically bind to a RSV antigen with a higher avidity than anypreviously known antibodies or fragments thereof.

[0100] The present invention also provides for antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens withan affinity constant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, atleast 5×10 M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹,at least 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least10¹² M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹,at least 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least5×10¹⁵ M⁻¹ and which have a higher avidity for one or more RSV antigensthan any previously known antibodies or fragments thereof such as, e.g.,SYNAGIS®. The present invention further provides pharmaceuticalcompositions comprising one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens with anaffinity constant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, atleast 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹²M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, atleast 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least5×10¹⁵ M⁻¹ and which have a higher avidity for one or more RSV antigensthan any previously known antibodies or fragments thereof such as, e.g.,SYNAGIS®.

[0101] The present invention provides methods of achieving a certainserum titer (preferably a serum titer 1 μg/ml or less, 2 μg/ml or less,5 μg/ml or less, 6 μg/ml or less, 10 μg/ml or less, 15 μg/ml or less, 20μg/ml or less, or 25 μg/ml or less) of antibodies or fragments thereofthat immunospecifically bind to one or more RSV antigens in a mammal,said methods comprising administering to said mammal one or moreantibodies or fragments thereof that have an affinity constant of atleast 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, atleast 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹²M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹ for said RSVantigens. Preferably, the antibodies or antibody fragments have a higheraffinity for a RSV antigen than SYNAGIS® does for the RSV Fglycoprotein.

[0102] The present invention also provides methods of achieving acertain serum titer of antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens in a mammal, saidmethods comprising administering to said mammal one or more antibodiesor fragments thereof that have a higher avidity for said RSV antigensthan any previously known antibodies or antibody fragments. Preferably,the antibodies or antibody fragments have higher avidity for a RSVantigen than SYNAGIS® has for the RSV F glycoprotein.

[0103] The present invention also provides methods of achieving acertain serum titer of antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens in mammal, saidmethods comprising administering to said mammal one or more antibodiesor fragments thereof that have an affinity constant of at least 2×10⁸M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, atleast 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, atleast 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹ for one or more RSVantigens and have a higher avidity than any previously known antibodiesor antibody fragments for said RSV antigens.

[0104] The present invention also provides methods of neutralizing RSVusing antibodies or fragments thereof that immunospecifically bind toone or more RSV antigens and which have an affinity constant of at least2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹,at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹,at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹ for said RSVantigens. Preferably, the antibodies or antibody fragments have a higheraffinity for a RSV antigen than SYNAGIS® does for the RSV Fglycoprotein. The present invention also provides methods ofneutralizing RSV using antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens and which have ahigher avidity for said RSV antigens than any previously knownantibodies or antibody fragments. Preferably, the antibodies or antibodyfragments have a higher avidity for a RSV antigen than SYNAGIS® does forthe RSV F glycoprotein. The present invention also provides methods ofneutralizing RSV using antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens with an affinityconstant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M, atleast 5×10¹² M⁻¹, at least 10¹³M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹ andwhich have a higher avidity for said RSV antigens than any previouslyknown antibodies or antibody fragments. The higher affinity and/orhigher avidity that these antibodies or antibody fragments have for aRSV antigen results in a lower concentration of these antibodies orantibody fragments necessary to achieve neutralization of RSV thanpreviously known.

[0105] The present invention also provides methods for preventing,treating or ameliorating one or more symptoms of RSV infection in amammal, said methods comprising administering to said mammal one or moreantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens and which have an affinity constant of at least 2×10⁸M⁻¹, at least 2.5×10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, atleast 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, atleast 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹ for said RSV antigens.Preferably, the antibodies or antibody fragments have a higher affinityfor a RSV antigen than SYNAGIS® does for the RSV F glycoprotein. Thepresent invention also provides of methods preventing, treating orameliorating one or more symptoms of RSV infection in a mammal, saidmethods comprising administering to said mammal one or more antibodiesor fragments thereof that immunospecifically bind to one or more RSVantigen and which have a higher avidity for said RSV antigen than anypreviously known antibodies or antibody fragments. Preferably, theantibodies or antibody fragments have a higher avidity for a RSV antigenthan SYNAGIS® does for the RSV F glycoprotein. The present inventionfurther provides methods of preventing, treating or ameliorating one ormore symptoms of RSV infection in a mammal, said methods comprisingadministering to said mammal one or more antibodies or fragments thereofthat immunospecifically bind to one or more RSV antigens with anaffinity constant of at least 2×10⁸ M⁻¹, at least 2.5×10⁸ M⁻¹, at least5×10⁸ M⁻¹, at least 10⁹ M⁻¹, at least 5×10⁹ M⁻¹, at least 10¹⁰ M⁻¹, atleast 5×10¹⁰ M⁻¹, at least 10¹¹ M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹²M⁻¹, at least 5×10¹² M⁻¹, at least 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, atleast 10¹⁴ M⁻¹, at least 5×10¹⁴ M⁻¹, at least 10¹⁵ M⁻¹, or at least5×10¹⁵ M⁻¹ for said RSV antigen and which have a higher avidity for saidRSV antigens than any previously known antibodies or antibody fragments.The higher affinity and/or higher avidity that these antibodies orantibody fragments have for a RSV antigen results in lower and/or lessfrequent doses of these antibodies or antibody fragments to achieve aprophylactic or therapeutic effect in a mammal, preferably a human, thanpreviously known.

[0106] The present invention provides methods for preventing, treating,or ameliorating one or more symptoms associated with a RSV infection ina mammal, preferably a human, said methods comprising administering afirst dose to said mammal of a prophylactically or therapeuticallyeffective amount of one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens with a higheravidity and/or affinity than previously known antibodies such asSYNAGIS®, wherein said effective amount is less than 15 mg/kg of saidantibodies or fragments thereof and which results in a serum titer lessthan 30 μg/ml (which is preferably at least 2 μg/ml, more preferably atleast 4 μg/ml, and most preferably at least 6 μg/ml) 30 days after thefirst administration and prior to any subsequent administration. In oneembodiment, RSV infection in a human subject is prevented or treated, orone or more symptoms in a human subject is ameliorated by administeringa first dose of less than 10 mg/kg, preferably less than 5 mg/kg, lessthan 3 mg/kg, less than 1 mg/kg, or less than 0.5 mg/kg of one or moreantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens with a higher avidity and/or affinity than previouslyknown antibodies such as SYNAGIS® so that a serum antibody titer of atleast 6 μg/ml, preferably at least 10 μg/ml, at least 25 μg/ml, at least30 μg/ml, at least 40 μg/ml at least 80 μg/ml, or at least 120 μg/ml, atleast 150 μg/ml, at least 200 μg/ml, at least 250 μ/ml, or at least 300μg/ml is achieved 30 days after the administration of the first dose ofthe antibodies or antibody fragments and prior to the administration ofa subsequent dose.

[0107] The present invention provides antibodies or fragments thereofcomprising a variable heavy (“VH”) domain having an amino acid sequenceof any VH domain listed in Table 2, and pharmaceutical compositionscomprising said antibodies or antibody fragments. The present inventionalso provides antibodies or fragments thereof comprising one or more VHCDRs having the amino acid sequence of one or more VH CDRs listed inTable 2 and/or Table 3, and pharmaceutical compositions comprising saidantibodies or antibody fragments. The present invention also providesantibodies or fragments thereof comprising a variable light (“VL”)domain having the amino acid sequence of any VL domain listed in Table2, and pharmaceutical compositions comprising said antibodies orantibody fragments. The present invention also provides antibodies orfragments thereof comprising one or more VL CDRs having the amino acidsequence of one or more VL CDRs listed in Table 2 and/or Table 3, andpharmaceutical compositions comprising said antibodies or antibodyfragments. The present invention also provides antibodies or fragmentsthereof comprising a VH domain having the amino acid sequence any VHdomain listed in Table 2 and a VL domain having the amino acid sequenceof any VL domain listed in Table 2, and pharmaceutical compositionscomprising said antibodies or antibody fragments. The present inventionprovides antibodies or fragments thereof comprising one or more VH CDRshaving the amino acid sequence one or more VH CDRs listed in Table 2and/or Table 3 and one or more VL CDRs having the amino acid sequence ofone or more VL CDRs listed in Table 2 and/or Table 3. The presentinvention encompasses pharmaceutical compositions comprising saidantibodies or antibody fragments. Preferably, said antibodies orantibody fragments immunospecifically bind to one or more RSV antigens.

[0108] The present invention encompasses methods for preventing,treating, neutralizing and ameliorating one or more symptoms using oneor more antibodies comprising a variable heavy (“VH”) domain having anamino acid sequence of any VH domain listed in Table 2. The presentinvention also encompasses methods for preventing, treating,neutralizing and ameliorating one or more symptoms using one or moreantibodies comprising one or more VH CDRs having the amino acid sequenceof one or more VH CDRs listed in Table 2 and/or Table 3. The presentinvention also encompasses methods for preventing, treating,neutralizing and ameliorating one or more symptoms using one or moreantibodies comprising a variable light (“VL”) domain having the aminoacid sequence of any VL domain listed in Table 2. The present inventionalso encompasses methods for preventing, treating, neutralizing andameliorating one or more symptoms using one or more antibodiescomprising one or more VL CDRs having the amino acid sequence of one ormore VL CDRs listed in Table 2 and/or Table 3. The present inventionalso encompasses methods for preventing, treating, neutralizing andameliorating one or more symptoms using one or more antibodiescomprising a VH domain having the amino acid sequence any VH domainlisted in Table 2 and a VL domain having the amino acid sequence of anyVL domain listed in Table 2. The present invention further encompassesmethods for preventing, treating, neutralizing and ameliorating one ormore symptoms using one or more antibodies comprising one or more VHCDRs having the amino acid sequence one or more VH CDRs listed in Table2 and/or Table 3 and one or more VL CDRs having the amino acid sequenceof one or more VL CDRs listed in Table 2 and/or Table 3. Preferably,said antibodies or antibody fragments immunospecifically bind to one ormore RSV antigens.

[0109] The present invention encompasses antibodies or fragments thereofthat immunospecifically bind to one or more RSV antigens with increasedin vivo half-lives. In particular, the present invention encompassesHL-SYNAGIS and antigen-binding fragments thereof. The present inventionalso encompasses novel antibodies or fragments thereof described hereinwhich immunospecifically bind to one or more RSV antigens and have an Fcdomain with a higher affinity for the FcRn receptor than the Fc domainof SYNAGIS®.

[0110] The present invention also encompasses methods for theprevention, neutralization, treatment or amelioration of one or moresymptoms associated with a RSV infection using antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens withincreased in vivo half-lives. In particular, the invention encompassesmethods for the prevention, neutralization, treatment or amelioration ofone or more symptoms associated with a RSV infection using HL-SYNAGIS oran antigen-binding fragment thereof The invention also encompassesmethods for the prevention, neutralization, treatment or amelioration ofone or more symptoms associated with a RSV infection using novelantibodies or fragments thereof described herein whichimmunospecifically bind to one or more RSV antigens and have an Fcdomain with a higher affinity for the FcRn receptor than the Fc domainof SYNAGIS®.

[0111] The present invention provides sustained release formulations ofantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens for the prevention, neutralization, treatment oramelioration of one or more symptoms associated with a RSV infection. Inparticular, the present invention provides sustained releaseformulations of SYNAGIS® or fragments thereof for the prevention,neutralization, treatment or amelioration of one or more symptomsassociated with a RSV infection. The present invention also providessustained release formulations of one or more novel antibodies orfragments thereof described herein which immunospecifically bind to oneor more RSV antigens for the prevention, neutralization, treatment oramelioration of one or more symptoms associated with a RSV infection.

[0112] The present invention also provides methods of administeringcompositions comprising antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens to the site of a RSVinfection in a subject. In particular, the present invention providescompositions comprising one or more antibodies or fragments thereof forpulmonary delivery to a subject.

[0113] The present invention provides compositions comprising one ormore antibodies or fragments thereof that immunospecifically bind to oneor more RSV antigens, and methods for detecting or diagnosing a RSVinfection utilizing said antibodies or antibody fragments.

[0114] 5.1. Antibodies

[0115] It should be recognized that antibodies that immunospecificallybind to a RSV antigen are known in the art. For example, SYNAGIS® is ahumanized monoclonal antibody presently used for the prevention of RSVinfection in pediatric patients. The present invention encompasses novelformulations for administration of SYNAGIS® and other known anti-RSVantibodies and novel doses of SYNAGIS® and other known anti-RSVantibodies, as discussed herein.

[0116] In addition, the invention encompasses novel antibodies,fragments and other biological or macromolecules whichimmunospecifically bind to one or more RSV antigens. With respect tothese novel agents, the invention further encompasses novel modes ofadministration, doses, dosing and uses based, in part, upon their uniquetherapeutic profiles and potency.

[0117] Set forth below, is a more detailed description of the antibodiesencompassed within the various aspects of the invention.

[0118] The present invention provides antibodies or fragments thereofthat immunospecifically bind to one or more RSV antigens. The presentinvention provides antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens. Preferably, theantibodies of the invention or fragments thereof immunospecifically bindto one or more RSV antigens regardless of the strain of RSV. The presentinvention also provides antibodies or fragments thereof thatdifferentially or preferentially bind to RSV antigens from one strain ofRSV versus another RSV strain. In a specific embodiment, the antibodiesof the invention or fragments thereof immunospecifically bind to the RSVF glycoprotein, G glycoprotein or SH protein. In a preferred embodiment,the antibodies present invention or fragments thereof immunospecificallybind to the RSV F glycoprotein. In another preferred embodiment, theantibodies of the present invention or fragments thereof bind to the A,B, or C antigenic sites of the RSV F glycoprotein.

[0119] Antibodies of the invention include, but are not limited to,monoclonal antibodies, multispecific antibodies, human antibodies,humanized antibodies, chimeric antibodies, single-chain Fvs (scFv),single chain antibodies, Fab fragments, F(ab′) fragments,disulfide-linked Fvs (sdFv), and anti-idiotypic (anti-Id) antibodies(including, e.g., anti-Id antibodies to antibodies of the invention),and epitope-binding fragments of any of the above. In particular,antibodies of the present invention include immunoglobulin molecules andimmunologically active portions of immunoglobulin molecules, i.e.,molecules that contain an antigen binding site that immunospecificallybinds to a RSV antigen. The immunoglobulin molecules of the inventioncan be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g.,IgG₁, IgG₂, IgG₃, IgG₄, IgA₁ and IgA₂) or subclass of immunoglobulinmolecule.

[0120] The antibodies of the invention may be from any animal originincluding birds and mammals (e.g., human, murine, donkey, sheep, rabbit,goat, guinea pig, camel, horse, or chicken). Preferably, the antibodiesof the invention are human or humanized monoclonal antibodies. As usedherein, “human” antibodies include antibodies having the amino acidsequence of a human immunoglobulin and include antibodies isolated fromhuman immunoglobulin libraries or from mice that express antibodies fromhuman genes.

[0121] The antibodies of the present invention may be monospecific,bispecific, trispecific or of greater multispecificity. Multispecificantibodies may be specific for different epitopes of a RSV polypeptideor may be specific for both a RSV polypeptide as well as for aheterologous epitope, such as a heterologous polypeptide or solidsupport material. See, e.g., PCT publications WO 93/17715, WO 92/08802,WO 91/00360, and WO 92/05793; 20 Tutt, et al., J. Immunol.147:60-69(1991); U.S. Pat. Nos. 4,474,893, 4,714,681, 4,925,648,5,573,920, and 5,601,819; and Kostelny et al., J. Immunol. 148:1547-1553(1992).

[0122] The present invention provides for antibodies or fragmentsthereof that exhibit a high potency in an assay described herein. Highpotency antibodies or fragments thereof can be produced by methodsdisclosed in copending U.S. patent application Ser. Nos. 60/168,426 and60/186,252 and methods described herein. For example, high potencyantibodies can be produced by genetically engineering appropriateantibody gene sequences and expressing the antibody sequences in asuitable host. The antibodies produced can be screened to identifyantibodies with, e.g., high k_(on) values in a BIAcore assay.

[0123] The present invention provides for antibodies or fragmentsthereof that have a high binding affinity for one or more RSV antigens.In a specific embodiment, an antibody of the present invention orfragment thereof has an association rate constant or

[0124] of at least 10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶ M⁻¹s⁻¹, at least 10⁷ M⁻¹s⁻¹, at least 5×10⁷M⁻¹s⁻¹, or at least 10⁸ M⁻¹s⁻¹. In a preferred embodiment, an antibodyof the present invention or fragment thereof has a k_(on) of at least2×10⁵ M⁻¹s⁻¹, at least 5×10⁵ M⁻¹s⁻¹, at least 10⁶M⁻¹s⁻¹, at least 5×10⁶M⁻¹s⁻¹, at least 10⁷M⁻¹ s⁻¹, at least 5×10⁷ M⁻¹ s⁻¹, or at least 10⁸M⁻¹s⁻¹.

[0125] In another embodiment, an antibody of the present invention orfragment thereof has a

[0126] of less than 10⁻¹s⁻¹, less than 5×10⁻¹ s⁻¹, less than 10² s⁻¹,less than 5×10⁻² s⁻¹, less than 10⁻³ s⁻¹, less than 5×10⁻³ s⁻¹, lessthan 10⁻⁴ s⁻¹, less than 5×10⁻⁴ s⁻¹, less than 10⁻⁵ s⁻¹, less than5×10⁻⁵ s⁻¹, less than 10⁻⁶ s⁻¹, less than 5×10⁻⁶ s⁻¹, less than 10⁻⁷s⁻¹, less than 5×10⁻⁷ s⁻¹, less than 10⁻⁸ s⁻¹, less than 5×10⁻⁸ s⁻¹,less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹ s⁻¹, or less than 10⁻¹⁰ s⁻¹. In apreferred embodiment, an antibody of the present invention or fragmentthereof has a k_(on) of less than 5×10⁻⁴ s⁻¹, less than 10⁻⁵ s⁻¹, lessthan 5×10⁻⁵ s⁻¹, less than 10⁻⁶ s⁻¹, less than 5×10⁻⁶ s⁻¹, less than10⁻⁷ s⁻¹, less than 5×10⁻⁷ s⁻¹, less than 10⁻⁸ s⁻¹, less than 5×10⁻⁸s⁻¹, less than 10⁻⁹ s⁻¹, less than 5×10⁻⁹ s⁻¹, or less than 10⁻¹⁰ s⁻¹

[0127] In another embodiment, an antibody of the present invention orfragment thereof has an affinity constant or K_(a)(k_(on)/k_(off)) of atleast 10² M⁻¹, at least 5×10² M⁻¹, at least 10³ M⁻¹, at least 5×10³ M⁻¹,at least 10⁴ M⁻¹, at least 5×10⁴ M⁻¹, at least 10⁵ M⁻¹, at least 5×10⁵M⁻¹, at least 10⁶ M⁻¹, at least 5×10⁶ M⁻¹, at least 10⁷ M⁻¹, at least5×10⁷ M⁻¹, at least 10⁸ M⁻¹, at least 5×10⁸ M⁻¹, at least 10⁹ M⁻¹, atleast 5×10 M⁻¹, at least 10¹⁰ M⁻¹, at least 5×10¹⁰ M⁻¹, at least 10¹¹M⁻¹, at least 5×10¹¹ M⁻¹, at least 10¹² M⁻¹, at least 5×10¹² M⁻¹, atleast 10¹³ M⁻¹, at least 5×10¹³ M⁻¹, at least 10¹⁴ M⁻¹, at least 5×10¹⁴M⁻¹, at least 10¹⁵ M⁻¹, or at least 5×10¹⁵ M⁻¹. In yet anotherembodiment, an antibody or fragment thereof has a dissociation constantor K_(d) (k_(off)/k_(on)) of less than 10⁻² M, less than 5×10⁻² M lessthan 10⁻³ M, less than 5×10⁻³ M, less than 10⁻⁴ M, less than 5×10⁻⁴ M,less than 10⁻⁵ M, less than 5×10⁻⁵ M, less than 10⁻⁶ M, less than 5×10⁻⁶M, less than 10⁻⁷ M, less than 5×10⁻⁷ M, less than 10⁻⁸ M, less than5×10⁻⁸ M, less than 10⁻⁹ M, less than 5×10⁻⁹ M, less than 10⁻¹⁰ M, lessthan 5×10⁻¹⁰ M, less than 10⁻¹¹ M, less than 5×10⁻¹¹ M, less than 10⁻¹²M, less than 5×10⁻¹² M, less than 10⁻¹³ M, less than 5×10⁻¹³ M, lessthan 10⁻¹⁴ M, less than 5×10⁻¹⁴ M, less than 10⁻¹⁵ M, or less than5×10⁻¹⁵ M.

[0128] The present invention provides antibodies or fragment thereofthat have a median effective concentration (EC₅₀) of less than 0.01 nM,less than 0.025 nM, less than 0.05 nM, less than 0.1 nM, less than 0.25nM, less than 0.5 nM, less than 0.75 nM, less than 1 nM, less than 1.25nM, less than 1.5 nM, less than 1.75 nM, or less than 2 nM, in an invitro microneutralization assay The median effective concentration isthe concentration of antibody or antibody fragments that neutralizes 50%of the RSV in an in vitro microneutralization assay. In a preferredembodiment, antibody of the invention or fragment thereof has an EC₅₀ ofless than 0.01 nM, less than 0.025 nM, less than 0.05 nM, less than 0.1nM, less than 0.25 nM, less than 0.5 nM, less than 0.75 nM, less than 1nM, less than 1.25 nM, less than 1.5 nM, less than 1.75 nM, or less than2 nM, in an in vitro microneutralization assay

[0129] In a specific embodiment, an antibody of the present invention isSYNAGIS® or an antibody-binding fragment thereof (e.g., one or morecomplementarity determining regions (CDRs) of SYNAGIS®). The amino acidsequence of SYNAGIS® is disclosed, e.g., in Johnson et al., 1997, J.Infectious Disease 176:1215-1224, and U.S. Pat. No. 5,824,307, each ofwhich is incorporated herein by reference in its entirety. Inalternative embodiment, an antibody of the present invention or fragmentthereof is not SYNAGIS® or a fragment of SYNAGIS®, i.e., is not anantibody comprising a VH domain of SEQ ID NO: 7 and/or a VL domain ofSEQ ID NO: 8.

[0130] The present invention provides for antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising the amino acid sequence ofSYNAGIS® with one or more amino acid residue substitutions in thevariable light (VL) domain and/or variable heavy (VH) domain depicted inFIG. 1. The present invention also provides for antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising the amino acid sequence ofSYNAGIS® with one or more amino acid residue substitutions in one ormore VL CDRs and/or one or more VH CDRs. In a specific embodiment, anantibody or fragment thereof comprises the amino acid sequence ofSYNAGIS® with one or more amino acid residue substitutions of the aminoacid residues indicated in bold face and underlining in Table 1. Inaccordance with this embodiment, the amino acid residue substitutionscan be conservative or non-conservative. The antibody or antibodyfragment generated by introducing substitutions in the VH domain, VHCDRs, VL domain and/or VL CDRs of SYNAGIS® can be tested in vitro and invivo, for example, for its ability to bind to RSV F antigen, for itsability to neutralize RSV, or for its ability to prevent, treat orameliorate one or more symptoms associated with a RSV infection. TABLE 1CDR Sequences Of SYNAGIS ® CDR Sequence SEQ ID NO: VH1 T S GMSVG 1 VH2DIWWD D K KD YNPSLK S 2 VH3 S MI T N W YFDV 3 VL1 KCQL SVGYMH 4 VL2 DTSKLA S 5 VL3 FQGS G YP F T 6

[0131] In a specific embodiment, an antibody of the present invention isSYNAGIS®, AFFF, Pf12, P12f4, P11d4, Ale9, A12a6, A13c4, A17d4, A4B4,A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, or A4B4-F52S.Preferably, an antibody of the present invention is AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9,Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S. In another embodiment, an antibody is a Fabfragment of SYNAGIS®, AFFF, Pf12, P12f4, P11d4, Ale9, A12a6, A13c4,A17d4, A4B4, A8c7, A4B4(1), A4B4L1FR-S28R, or A4B4-F52S.

[0132] AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A8C7,1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10,A13a11, and A1h5 comprise the framework region and constant regions ofSYNAGIS®. A4B4, A4B4(1), A4B4L1FR-S28R, and A4B4-F52S comprise theframework region and constant regions of SYNAGIS® with the exceptionthat there is an amino acid substitution of an alanine for a valine atposition 109. In certain embodiments, A4B4, A4B4(1), A4B4L1FR-S28R, andA4B4-F52S comprise the framework region and constant regions ofSYNAGIS®.

[0133] In another embodiment, the present invention provides for anantigen-binding fragment of AFFF, Pf12, P12f4, P11d4, A1e9, A12a6,A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1),6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, orA4B4-F52S. In another embodiment, the present invention provides for oneor more antibodies or fragments thereof that immunospecifically bind toone or more RSV antigens, said antibodies or antibody fragmentscomprising a VH chain and/or VL chain having the amino acid sequence ofa VH chain and/or VL chain of AFFF, Pf12, P12f4, P11d4, A1e9, A12a6,A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1),6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, orA4B4-F52S. In another embodiment, the present invention provides for oneor more antibodies or fragments thereof that immunospecifically bind toone or more RSV antigens, said antibodies or antibody fragmentscomprising a VH domain and/or VL domain having the amino acid sequenceof a VH domain and/or VL domain of AFFF, Pf12, P12f4, P11d4, A1e9,A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG,AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, orA4B4-F52S. In another embodiment, the present invention provides forantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens, said antibodies or antibody fragments comprising oneor more CDRs having the amino acid sequence of one or more CDRs of AFFF,Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8c7, 1X-493L1FR,H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5,A4B4(1), A4B4L1FR-S28R, or A4B4-F52S. In yet another embodiment, thepresent invention provides for one or more antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising a combination of VH CDRsand/or VL CDRs having the amino acid sequence of VH CDRs and/or VL CDRsof AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8c7,1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10,A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, and/or A4B4-F52S.

[0134] The present invention provides antibodies or fragments thereofthat immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising a variable heavy (“VH”)chain having an amino acid sequence of any one of the VL domains listedin Table 2. The present invention also provides antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising a VH domain having an aminoacid sequence of any one of the VH domains listed in Table 2. In certainembodiments of the invention, an antibody or fragment thereof comprisinga VH domain having an amino acid of any of one of the VH domains listedin Table 2 is not SYNAGIS®. The present invention also providesantibodies or fragments thereof that immunospecifically bind to one ormore RSV antigens, said antibodies or fragments comprising a VHcomplementarity determining region (“CDR”) having an amino acid sequenceof any one of the VH CDRs listed in Table 2 and/or Table 3. In certainembodiments of the invention, an antibody or fragment thereof comprisinga VH CDR having an amino acid of any of one of the VH CDRs listed inTable 2 and/or Table 3 is not SYNAGIS®. TABLE 2. Antibodies & FragmentsThereof Antibody VH VH VL VL Name Chain Domain VH CDR1 VH CDH2 VH CDR3Chain Domain VL CDR1 VL CDR2 VL CDR3 **SYNAGIS SEQ SEQ ID TSGMSVGDIWWDDKKDY SMITNWYFDV SEQ SEQ ID NO:8 KCQLSVGYM DTSKLAS FQGSGYPFT ID NO:NO:7 (SEQ ID NPSLKS (SEQ ID ID NO: H (SEQ ID (SEQ ID 208 NO:1) (SEQ IDNO:3) 209 (SEQ ID NO:5) NO:6) NO:2) NO:4) ***AFFF SEQ SEQ ID T A GMSVGDIWWDDKKDYN SMITN F YFDV SEQ SEQ ID NO:13 SASS SVGYMH DT F KLAS FQ FSGYPFT ID NO: NO:9 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID210 NO:10) (SEQ ID NO:12) 211 NO:14) NO:15) NO:16) NO:2) ***P12f2 SEQSEQ ID TGMSVG DIWWDDKK H YN D MI F N F YFDV SEQ SEQ ID NO:21 SLSSRRVGYMH DT FY L S S FQGSGYPFT ID NO: NO:17 (SEQ ID PSLK D (SEQ ID ID NO:(SEQ ID (SEQ ID (SEQ ID 212 NO:18) (SEQ ID NO:20) 213 NO:22) NO:23)NO:6) NO:19) ***P12f4 SEQ SEQ ID T PGMSVG DIWWD G KK H YN D MI F N FYFDV SEQ SEQ ID NO:26 SLSSR VGYMH DT RG L P S FQGSGYPFT ID NO: NO:24(SEQ ID PSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 214 NO:18) (SEQ IDNO:20) 215 NO:22) NO:27) NO:6) NO:25) ***P11d4 SEQ SEQ ID T P GMSVGDIWWD G KK H YN D MI F NWYFDV SEQ SEQ ID NO:30 SPSSR VGYMH DT MR LASFQGSGYPFT ID NO: NO:28 (SEQ ID PSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID(SEQ ID 216 NO:18) (SEQ ID NO:29) 217 NO:31) NO:32) NO:6) NO:25) ***Ale9SEQ SEQ ID T A GMSVG DIWWD G KK H YN D MI F NWYFDV SEQ SEQ ID NO:34SLSSR VGYMH DT F KL S S FQGSGYPFT ID NO: NO:33 (SEQ ID PSLK D (SEQ ID IDNO: (SEQ ID (SEQ ID (SEQ ID 218 NO:10) (SEQ ID NO:29) 219 NO:22) NO:35)NO:6) NO:25) ***A12a6 SEQ SEQ ID T A GMSVG DIWWD G KKDYN D MI F N F YFDVSEQ SEQ ID NO:38 SASSR VGYMH DT F KL S S FQGSGYPFT ID NO: NO:36 (SEQ IDPSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 220 NO:10) (SEQ ID NO:20)221 NO:35) NO:6) NO:37) ***A13c4 SEQ SEQ ID T A GMSVG DIWWD G KK S YN DMI F N F YFDV SEQ SEQ ID NO:42 SLSSR VGYMH DT MYQ SS FQGSGYPFT ID NO:NO:40 (SEQ ID PSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 222 NO:10)(SEQ ID NO:20) 223 NO:22) NO:43) NO:6) NO:41) ***A17d4 SEQ SEQ ID T AGMSVG DIWWDDKK S Y D MI F N F YFDV SEQ SEQ ID NO:46 LPSSR VGYM DT MYQS SFQGSGYPFT ID NO: NO:44 (SEQ ID NPSLK D (SEQ ID ID NO: H (SEQ ID (SEQ ID224 NO:10) (SEQ ID NO:20) 225 (SEQ ID NO:43) NO:6) NO:45) NO:47) ***A4B4SEQ SEQ ID T A GMSVG DIWWDDKK H D MI F N F YFDV SEQ SEQ ID NO:49 SASSRVGYM DT FF L D S FQGSGYPFT ID NO: NO:48 (SEQ ID YNPSLK D (SEQ ID ID NO:H (SEQ ID (SEQ ID 226 NO:10) (SEQ ID NO:20) 227 (SEQ ID NO:50) NO:6)NO:19) NO:39) ***A8c7 SEQ SEQ ID T A GMSVG DIWWDDKK S Y D MI F NWYFD SEQSEQ ID NO:52 SPSSR VGYM DT RYQS S FQGSGYPFT ID NO: NO:51 (SEQ ID NPSLK DV ID NO: H (SEQ ID (SEQ ID 228 NO:10) (SEQ ID (SEQ ID 229 (SEQ ID NO:53)NO:6) NO:45) NO:29) NO:31) *1X- SEQ SEQ ID TSGMSVG DIWWDDKKDYNSMITNWYFDV SEQ SEQ ID NO:54 SASS SVGYMH DTSKLAS FQGSGYPFT 493L1FR ID NO:NO:7 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 230 NO:1) (SEQID NO:3) 231 NO:14) NO:5) NO:6) NO:2) *H3-3F4 SEQ SEQ ID T A GMSVGDIWWDDKKDYN D MI F NWYFDV SEQ SEQ ID NO:56 SASS SVGYMH DT F KLASFQGSGYPFT ID NO: NO:55 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQID 232 NO:10) (SEQ ID NO:29) 233 NO:14) NO:15) NO:6) NO:2) *M3H9 SEQ SEQID T A GMSVG DIWWDDKKDYN D MI F NWYFDV SEQ SEQ ID NO:70 SASS SVGYMH DT YK QT S FQGSGYPFT ID NO: NO:55 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQID (SEQ ID 234 NO:10) (SEQ ID NO:29) 235 NO:14) NO:57) NO:6) NO:2)*Y10H6 SEQ SEQ ID T A GMSVG DIWWDDKKDYN DMIF NWYFDV SEQ SEQ ID NO:58SASS SVGYMH DT RY L S S FQGSGYPFT ID NO: NO:55 (SEQ ID PSLKS (SEQ ID IDNO: (SEQ ID (SEQ ID (SEQ ID 236 NO:10) (SEQ ID NO:29) 237 NO:14) NO:59)NO:6) NO:2) *DG SEQ SEQ ID T A GMSVG DIWWDDKKDYN D MITN F YFDV SEQ SEQID NO:56 SASS SVGYMH DT F KLAS FQGSGYPFT ID NO: NO:78 (SEQ ID PSLKS (SEQID ID NO: (SEQ ID (SEQ ID (SEQ ID 238 NO:10) (SEQ ID NO:79) 239 NO:14)NO:15) NO:6) NO:2) AFFF(1) SEQ SEQ ID T A GMSVG DIWWDDKKDYN SMITNF YFDVSEQ SEQ ID NO:60 SASS SVGYMH DT F KLAS FQGS F YPFT ID NO: NO:9 (SEQ IDPSLKS (SEQ ID (SEQ ID (SEQ ID (SEQ ID 240 NO:10) (SEQ ID NO:12) 241NO:14) NO:15) NO:61) NO:2) *6H8 SEQ SEQ ID T A GMSVG DIWWDDKKDYN D MITNF YFDV SEQ SEQ ID NO:62 SASS SVGYMH DT F KL T S FQGSGYPFT ID NO: NO:78(SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 242 NO:10) (SEQ IDNO:79) 243 NO:14) NO:63) NO:6) NO:2) *L1-7E5 SEQ SEQ ID T A GMSVGDIWWDDKKDYN D MITN F YFDV SEQ SEQ ID NO:64 SASSR VGYMH DT F KLASFQGSGYPFT ID NO: NO:78 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQID 244 NO:10) (SEQ ID NO:79) 245 NO:39) NO:15) NO:6) NO:2) *L2-45B10 SEQSEQ ID T A GMSVG DIWWDDKKDYN D MITN F YFDV SEQ SEQ ID NO:65 SASS SVGYMHDT FR LAS FQGSGYPFT ID NO: NO:78 (SEQ ID PSLKS (SEQ ID ID NO: (SEQ ID(SEQ ID (SEQ ID 246 NO:10) (SEQ ID NO:79) 247 NO:14) NO:66) NO:6) NO:2)*A13a11 SEQ SEQ ID T A GMSVG DIWWDDKK H YN D MI F NWYFDV SEQ SEQ IDNO:68 SPSSR VGYMH DT YRHS S FQGSGYPFT ID NO: NO:67 (SEQ ID PSLK D (SEQID ID NO: (SEQ ID (SEQ ID (SEQ ID 248 NO:10) (SEQ ID NO:29) 249 NO:31)NO:69) NO:6) NO:19) *A1h5 SEQ SEQ ID T AGMSVG DIWWD G KK H YN D MI FNWYFDV SEQ SEQ ID NO:71 SLSSS VGYMH DT FFHR S FQGSGYPFT ID NO: NO:33(SEQ ID PSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 250 NO:10) (SEQ IDNO:29) 251 NO:72) (NO:73) NO:6) NO:25) A4B4(1) SEQ SEQ ID T A GMSVGDIWWDDKK H D MI F N F YFDV SEQ SEQ ID NO:74 SASSR VGYM DT LL L D SFQGSGYPFT ID NO: NO:48 (SEQ ID YNPSLK D (SEQ ID ID NO: H (SEQ ID (SEQ ID252 NO:10) (SEQ ID NO:20) 253 (SEQ ID NO:75) NO:6) NO:20) (NO:39)***A4B4L1 SEQ SEQ ID T A GMSVG DIWWDDKK H D MI F N F YFDV SEQ SEQ IDNO:11 SASSR VGYM DTSKLAS FQGSGYPFT FR-S28R ID NO: NO:48 (SEQ ID YNPSLK D(SEQ ID ID NO: H (SEQ ID (SEQ ID 254 NO:10) (SEQ ID NO:20) 255 (SEQ IDNO:5) NO:6) NO:19) NO:39) ***A4B4 SEQ SEQ ID T A GMSVG DIWWDDKK H YN DMI F N F YFDV SEQ SEQ ID NO:76 SASSR VGYMH DTS F L D S FQGSGYPFT F52S IDNO: NO:48 (SEQ ID PSLK D (SEQ ID ID NO: (SEQ ID (SEQ ID (SEQ ID 256NO:10) (SEQ ID NO:20) 257 NO:39) NO:77) NO:6) NO:19)

[0135] TABLE 3 CDR Sequences VH CDR1 VH CDR2 VH CDR3 VL CDR1 VL CDR2 VLCDR3 T S GMSVG DIWWD D KK D YNPSLK S S MI T N W YFDV KCOLS VGYMH DT SK LA S FQGSGYPFT (SEQ ID NO:1) (SEQ ID NO:2) (SEQ ID NO:3) (SEQ ID NO:4)(SEQ ID NO:5) (SEQ ID NO:6) T P GMSVG DIWWDDKK H YNPSLK D D MI T N FYFDV KCOSSVGYMH DT SY L A S FQ F SGYPFT (SEQ ID NO:18) (SEQ ID NO:19)(SEQ ID NO:29) (SEQ ID NO 80) (SEQ ID NO:81) (SEQ ID NO:6) T A GMSVGDIWWDDKK H YNPSLK S D MI T N W YFDV KCOSR VGYMH DT SY L S S FQGS F YPFT(SEQ ID NO:10) (SEQ ID NO:82) (SEQ ID NO:83) (SEQ ID NO:84) (SEQ IDNO:85) (SEQ ID NO:61) DIWWD D KK D YNPSLK D D MI F N W YFDV KCQLR VGYMHDT KK L S S (SEQ ID NO:86) (SEQ ID NO:29) (SEQ ID NO:87) (SEQ ID NO:88)DIWWD D KK H YNPSLK S D MI F N F YFDV KLQLS VGYMH DT FY L S S (SEQ IDNO:91) (SEQ ID NO:20) (SEQ ID NO:89) (SEQ ID NO:90) DIWWD DKKD YNPSLK DS MI T N F YFDV KLQSS VGYMH DT FK L A S (SEQ ID NO:93) (SEQ ID NO:12)(SEQ ID NO:92) (SEQ ID NO:15) DIWWD G KK H YNPSLK D S MI F N W YFDVKLQSR VGYMH DT FK L S S (SEQ ID NO:25) (SEQ ID NO:94) (SEQ ID NO:95)(SEQ ID NO:96) DIWWD G KK D YNPSLK S S MI F N F YFDV KLQLR VGYMH DT FY LA S (SEQ ID NO:100) (SEQ ID NO:97) (SEQ ID NO:98) (SEQ ID NO:99) DIWWD GKK D YNPSLK D KLSLS VGYMH DT SK L P S (SEQ ID NO:103) (SEQ ID NO:101)(SEQ ID NO:102) DIWWD G KK H YNPSLK S KLSSS VGYMH DT SG L A S (SEQ IDNO:106) (SEQ ID NO:104) (SEQ ID NO:105) DIWWD D KK S YNPSLK S KLSSRVGYMH DT SG L P S** (SEQ ID NO:109) (SEQ ID NO:107) (SEQ ID NO:108)DIWWD D KK S YNPSLK D KLSLR VGYMH DT RG L P S (SEQ ID NO:111) (SEQ ID NO110) (SEQ ID NO:27) DIWWD G KK S YNPSLK S KCSLS VGYMH DT RK L A S (SEQID NO:114) (SEQ ID NO: 112) (SEQ ID NO:113) DIWWD G KK S YNPSLK D KCSSSVGYMH DT RG L A S (SEQ ID NO:41) (SEQ ID NO:115) (SEQ ID NO:116) KCSSRVGYMH DT RK L P S (SEQ ID NO:117) (SEQ ID NO:118) KCSLR VGYMH DT MR L AS (SEQ ID NO:119) (SEQ ID NO:32) SLSLS VGYMH DT MK L A S (SEQ ID NO:120)(SEQ ID NO:121) SLSSS VGYMH DT SR L A S (SEQ ID NO:122) (SEQ ID NO:123)SLSSR VGYMH DT SL L A S (SEQ ID NO:22) (SEQ ID NO:124) SLSSR VGYMH DT SLL D S (SEQ ID NO:125) (SEQ ID NO:126) SCQLS VGYMH DT SK L D S (SEQ IDNO:127) (SEQ ID NO:128) SCQSS VGYMH DT LL L D S (SEQ ID NO:129) (SEQ IDNO:75) SCQSR VGYMH DT LK L D S (SEQ ID NO:130) (SEQ ID NO:131) SCQLRVGYMH DT LL L A S (SEQ ID NO:132) (SEQ ID NO:133) SLQLSVGYMH DT LKLA S(SEQ ID NO:134) (SEQ ID NO:135) SLQSS VGYMH DT SKLS S (SEQ ID NO:136)(SEQ ID NO:137) SLQSR VGYMH DT SKQA S (SEQ ID NO:138) (SEQ ID NO:139)SLQLR VGYMH DT SKQS S (SEQ ID NO:140) (SEQ ID NO:141) SCSLS VGYMH DTSYLA S (SEQ ID NO:142) (SEQ ID NO:143) SCSSS VGYMH DT SYLS S (SEQ IDNO:144) (SEQ ID NO:145) SCSSR VGYMH DTSYQAS (SEQ ID NO:146) (SEQ IDNO:147) SCSLR VGYMH DT SYQS S (SEQ ID NO:148) (SEQ ID NO:149) KPSSRVGYMH DT MYQA S (SEQ ID NO:150) (SEQ ID NO:151) KPSLR VGYMH DT MYQS S(SEQ ID NO:152) (SEQ ID NO:143) KPSSS VGYMH DT MKQA S (SEQ ID NO:153)(SEQ ID NO:154) KPSLS VGYMH DT SKQS S (SEQ ID NO:155) (SEQ ID NO:156)KPQSR VGYMH DT MYLA S (SEQ ID NO:157) (SEQ ID NO:158) KPQLR VGYMH DTMYLS S (SEQ ID NO:159) (SEQ ID NO:160) KPQSS VGYMH DT MKLA S (SEQ IDNO:161) (SEQ ID NO:162) KPQLS VGYMH DT MKLS S (SEQ ID NO:163) (SEQ IDNO:164) SPSSRVGYMH DT SK K S S (SEQ ID NO:31) (SEQ ID NO:165) SPSLRVGYMH DT RYQA S (SEQ ID NO:166) (SEQ ID NO:167) SPSSS VGYMH DT RYQS S(SEQ ID NO:168) (SEQ ID NO:53) SPSLS VGYMH DT RKQA S (SEQ ID NO:169)(SEQ ID NO:170) SPQSR VGYMH DT RKQS S (SEQ ID NO:171) (SEQ ID NO:172)SPQLR VGYMH DT RKLA S (SEQ ID NO:173) (SEQ ID NO:174) SPQSS VGYMH DTRKLS S (SEQ ID NO:175) (SEQ ID NO:176) SPQLS VGYMH DT RYLA S (SEQ IDNO:177) (SEQ ID NO:178) KAQSR VGYMH DT RYLS S (SEQ ID NO:179) (SEQ IDNO:59) KAQLR VGYMH DT FF L D S (SEQ ID NO:180) (SEQ ID NO:50) KAQSSVGYMH DTS F L D S (SEQ ID NO:181) (SEQ ID NO:77) KAQLS VGYMH (SEQ IDNO:182) KASSR VGYMH (SEQ ID NO:183) KASLR VGYMH (SEQ ID NO:184) KASSSVGYMH (SEQ ID NO:185) KASLS VGYMH (SEQ ID NO:186) SASSR VGYMH (SEQ IDNO:39) SASLR VGYMH (SEQ ID NO:187) SASSS VGYMH (SEQ ID NO:14) SASLSVGYMH (SEQ ID NO:188) SAQSR VGYMH (SEQ ID NO:189) SAQLR VGYMH (SEQ IDNO:190) SAQSS VGYMH (SEQ ID NO:191) LPSSR VGYMH (SEQ ID NO:47) LPSLSVGYMH (SEQ ID NO:192) LPSSS VGYMH (SEQ ID NO:193) LPSLR VGYMH (SEQ IDNO:194) LCSSR VGYMH (SEQ ID NO:195) LCSLS VGYMH (SEQ ID NO:196) LCSSSVGYMH (SEQ ID NO:197) LCSLR VGYMH (SEQ ID NO:198) LPQSR VGYMH (SEQ IDNO:199) LPQLS VGYMH (SEQ ID NO:200) LPQSS VGYMH (SEQ ID NO:201) LPQLRVGYMH (SEQ ID NO:202) LCQSR VGYMH (SEQ ID NO:203) LCQLS VGYMH (SEQ IDNO:204) LCQSS VGYMH (SEQ ID NO:205) LCQLR VGYMH (SEQ ID NO:206) SAQLSVGYMH (SEQ ID NO:207)

[0136] In one embodiment of the present invention, antibodies orfragments thereof comprise a VH CDR1 having the amino acid sequence ofSEQ ID NO: 1, SEQ ID NO: 10 or SEQ ID NO: 18. In another embodiment,antibodies or fragments thereof comprise a VH CDR2 having the amino acidsequence of SEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 37,SEQ ID NO: 41 or SEQ ID NO: 45. In another embodiment, antibodiescomprise a VH CDR3 having the amino acid sequence of SEQ ID NO: 3, SEQID NO: 12, SEQ ID NO: 20, SEQ ID NO: 29 or SEQ ID NO: 79. In a preferredembodiment, antibodies or fragments thereof comprise a VH CDR1 havingthe amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 10 or SEQ ID NO: 18,a VH CDR2 having the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 19,SEQ ID NO: 25, SEQ ID NO: 37 or SEQ ID NO: 41, SEQ ID NO: 45, and a VHCDR3 having the amino acid sequence of SEQ ID NO: 3, SEQ ID NO: 12, SEQID NO: 20, SEQ ID NO: 29 or SEQ ID NO: 79.

[0137] The present invention also provides antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising a variable light (“VL”)domain having an amino acid sequence of any one of the VL domains listedin Table 2. The present invention also provides antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or fragments comprising a VL CDR having an amino acidsequence of any one of the VL CDRs listed in Table 2 and/or Table 3.

[0138] In one embodiment of the present invention, antibodies orfragments thereof comprise a VL CDR1 having the amino acid sequence ofSEQ ID NO: 4, SEQ ID NO: 14, SEQ ID NO: 22, SEQ ID NO: 31, SEQ ID NO:39, or SEQ ID NO: 47. In another embodiment, antibodies or fragmentsthereof comprise a VL CDR2 having the amino acid sequence of SEQ ID NO:5, SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 27, SEQ ID NO: 32, SEQ IDNO: 35, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO: 57, SEQID NO: 59, SEQ ID NO: 63, SEQ ID NO: 66, SEQ ID NO: 69, SEQ ID NO: 73,SEQ ID NO: 75 or SEQ ID NO: 77. In another embodiment, antibodies orfragments thereof comprise a VL CDR3 having the amino acid sequence ofSEQ ID NO: 6, SEQ ID NO: 16 or SEQ ID NO: 61. In a preferred embodiment,antibodies or fragments thereof comprise a VL CDR1 having the amino acidsequence of SEQ ID NO: 4, SEQ ID NO: 14, SEQ ID NO: 22, SEQ ID NO: 31,SEQ ID NO: 39, or SEQ ID NO: 47, a VL CDR2 having the amino acidsequence of SEQ ID NO: 5, SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 27,SEQ ID NO: 32, SEQ ID NO: 35, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO:53, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 63, SEQ ID NO: 66, SEQ IDNO: 69, SEQ ID NO: 73, SEQ ID NO: 75 or SEQ ID NO: 77, and a VL CDR3having the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 16 or SEQ IDNO: 61.

[0139] The present invention also provides antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising a VH domain disclosed hereincombined with a VL domain disclosed herein, or other VL domain. Thepresent invention further provides antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens, said antibodies orfragments comprising a VL domain disclosed herein combined with a VHdomain disclosed herein, or other VH domain. In a preferred embodiment,antibodies or fragments thereof that immunospecifically bind to a RSVantigen comprise a VH domain having the amino acid sequence of SEQ IDNO: 7, SEQ ID NO: 9, SEQ ID NO: 17, SEQ ID NO: 24, SEQ ID NO: 28, SEQ IDNO: 33, SEQ ID NO: 36, SEQ ID NO: 40, SEQ ID NO: 44, SEQ ID NO: 48, SEQID NO: 51, SEQ ID NO: 55, SEQ ID NO: 67 or SEQ ID NO: 78 and a VL domainhaving the amino acid sequence of SEQ ID NO: 8, SEQ ID NO: 13, SEQ IDNO: 21, SEQ ID NO: 26, SEQ ID NO: 30, SEQ ID NO: 34, SEQ ID NO: 38, SEQID NO: 42, SEQ ID NO: 46, SEQ ID NO: 49, SEQ ID NO: 52, SEQ ID NO: 54 ,SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO:64, SEQ ID NO: 65, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ IDNO: 74 or SEQ ID NO: 76.

[0140] The present invention also provides antibodies or fragmentsthereof comprising one or more VH CDRs and one or more VL CDRs listed inTable 2 and/or Table 3. In particular, the invention provides for anantibody or fragment thereof comprising a VH CDR1 and a VL CDR1, a VHCDR1 and a VL CDR2, a VH CDR1 and a VL CDR3, a VH CDR2 and a VL CDR1, VHCDR2 and VL CDR2, a VH CDR2 and a VL CDR3, a VH CDR3 and a VH CDR1, a VHCDR3 and a VL CDR2, a VH CDR3 and a VL CDR3, or any combination thereofof the VH CDRs and VL CDRs listed in Table 2. The invention alsoprovides for an antibody or fragment thereof comprising a VH CDR1 and aVL CDR1, a VH CDR1 and a VL CDR2, a VH CDR1 and a VL CDR3, a VH CDR2 anda VL CDR1, VH CDR2 and VL CDR2, a VH CDR2 and a VL CDR3, a VH CDR3 and aVH CDR1, a VH CDR3 and a VL CDR2, a VH CDR3 and a VL CDR3, or anycombination thereof of the VH CDRs and VL CDRs listed in Table 3. Theinvention also provides for an antibody or fragment thereof comprising aVH CDR1 and a VL CDR1, a VH CDR1 and a VL CDR2, a VH CDR1 and a VL CDR3,a VH CDR2 and a VL CDRl, VH CDR2 and VL CDR2, a VH CDR2 and a VL CDR3, aVH CDR3 and a VH CDR1, a VH CDR3 and a VL CDR2, a VH CDR3 and a VL CDR3,or any combination thereof of the VH CDRs and VL CDRs listed in Table 2and Table 3.

[0141] In one embodiment, an antibody or fragment thereof comprises a VHCDR1 having the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 10 orSEQ ID NO: 18 and a VL CDR1 having the amino acid sequence of SEQ ID NO:4, SEQ ID NO: 14, SEQ ID NO: 22, SEQ ID NO: 3 1, SEQ ID NO: 39, or SEQID NO: 47. In another embodiment, an antibody of the present inventionor fragment thereof comprises a VH CDR1 having the amino acid sequenceof SEQ ID NO: 1, SEQ ID NO: 10 or SEQ ID NO: 18 and a VL CDR2 having theamino acid sequence of SEQ ID NO: 5, SEQ ID NO: 15, SEQ ID NO: 23, SEQID NO: 27, SEQ ID NO: 32, SEQ ID NO: 35, SEQ ID NO: 43, SEQ ID NO: 50,SEQ ID NO: 53, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 63, SEQ ID NO:66, SEQ ID NO: 69, SEQ ID NO: 73, SEQ ID NO: 75, or SEQ ID NO: 77. Inanother embodiment, an antibody of the present invention or fragmentthereof comprises a VH CDR1 having the amino acid sequence of SEQ ID NO:1, SEQ ID NO: 10 or SEQ ID NO: 18 and a VL CDR3 having the amino acidsequence of SEQ ID NO: 6, SEQ ID NO: 16 or SEQ ID NO: 61.

[0142] In another embodiment, an antibody of the present invention orfragment thereof comprises a VH CDR2 having the amino acid sequence ofSEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 37, SEQ ID NO: 41or SEQ ID NO: 45 and a VL CDR1 having the amino acid sequence of SEQ IDNO: 4, SEQ ID NO: 14, SEQ ID NO: 22, SEQ ID NO: 31, SEQ ID NO: 39 or SEQID NO: 47. In another embodiment, an antibody of the present inventionor fragment thereof comprises a VH CDR2 having the amino acid sequenceof SEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 37, SEQ ID NO:41 or SEQ ID NO: 45 and a VL CDR2 having the amino acid sequence of SEQID NO: 5, SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 27, SEQ ID NO: 32,SEQ ID NO: 35, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO:57, SEQ ID NO: 59, SEQ ID NO: 63, SEQ ID NO: 66, SEQ ID NO: 69, SEQ IDNO: 73, SEQ ID NO: 75, or SEQ ID NO: 77. In another embodiment, anantibody of the present invention or fragment thereof comprises a VHCDR2 having the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 19, SEQID NO: 25, SEQ ID NO: 37, SEQ ID NO: 41 or SEQ ID NO: 45 and a VL CDR3having the amino acid sequence of SEQ ID NO: 6, SEQ ID NO: 16, or SEQ IDNO: 61.

[0143] In another embodiment, an antibody of the present invention orfragment thereof comprises a VH CDR3 having the amino acid sequence ofSEQ ID NO: 3, SEQ ID NO: 12, SEQ ID NO: 20, SEQ ID NO: 29 or SEQ ID NO:79 and a VL CDR1 having the amino acid sequence of SEQ ID NO: 4, SEQ IDNO: 14, SEQ ID NO: 22, SEQ ID NO: 31, SEQ ID NO: 39 or SEQ ID NO: 47. Inanother embodiment, an antibody of the present invention or fragmentthereof comprises a VH CDR3 having the amino acid sequence of SEQ ID NO:3, SEQ ID NO: 12, SEQ ID NO: 20, SEQ ID NO: 29 or SEQ ID NO: 79 and a VLCDR having the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 15, SEQID NO: 23, SEQ ID NO: 27, SEQ ID NO: 32, SEQ ID NO: 35, SEQ ID NO: 43,SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO:63, SEQ ID NO: 66, SEQ ID NO: 69, SEQ ID NO: 73, SEQ ID NO: 75, or SEQID NO: 77. In a preferred embodiment, an antibody of the presentinvention or fragment thereof comprises a VH CDR3 having the amino acidsequence of SEQ ID NO: 3, SEQ ID NO: 12, SEQ ID NO: 20, SEQ ID NO: 29 orSEQ ID NO: 79 and a VL CDR3 having the amino acid sequence of SEQ ID NO:6, SEQ ID NO: 16, or SEQ ID NO: 61.

[0144] The present invention also provides for nucleic acid molecules,generally 10 isolated, encoding an antibody of the invention or fragmentthereof. In a specific embodiment, isolated nucleic acid molecules ofthe invention encode for SYNAGIS®, AFFF, Pf12, P12f4, P11d4, A1e9,A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG,AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, orA4B4-F52S. Preferably, isolated nucleic acid molecules of the inventionencode for AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4,A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, or A4B4-F52S. In anotherembodiment, isolated nucleic acid molecules of the invention encode foran antigen-binding fragment of SYNAGIS®, AFFF, Pf12, P12f4, P11d4, A1e9,A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG,AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, orA4B4-F52S.

[0145] In another embodiment, an isolated nucleic acid molecule(s) ofthe invention encodes an antibody or fragment thereof comprising a VHdomain having an amino acid sequence of any one of the VH domains listedin Table 2. In another embodiment, an isolated nucleic acid molecule(s)of the invention encodes an antibody or fragment thereof comprising a VHCDR1 having an amino acid sequence of any one of the VH CDR1s listed inTable 2 or Table 3. In another embodiment, an isolated nucleic acidmolecule(s) of the invention encodes an antibody or fragment thereofcomprising a VH CDR2 having an amino acid sequence of any one of the VHCDR2s listed in Table 2 or Table 3. In yet another embodiment, anisolated nucleic acid molecule(s) of the invention encodes an antibodyor fragment thereof comprising a VH CDR3 having an amino acid sequenceof any one of the VH CDR3s listed in Table 2 or Table 3.

[0146] In another embodiment, an isolated nucleic acid molecule(s) ofthe invention encodes an antibody or fragment thereof comprising a VLdomain having an amino acid sequence of any one of the VL domains listedin Table 2. In another embodiment, an isolated nucleic acid molecule(s)of the present invention encodes an antibody or fragment thereofcomprising a VL CDR1 having amino acid sequence of any one of the VLCDR1s listed in Table 2 or Table 3. In another embodiment, an isolatednucleic acid molecule(s) of the present invention encodes an antibody orfragment thereof comprising a VL CDR2 having an amino acid sequence ofany one of the VL CDR2s listed in Table 2 or Table 3. In yet anotherembodiment, an isolated nucleic acid molecule(s) of the presentinvention encodes an antibody or fragment thereof comprising a VL CDR3having an amino acid sequence of any one of the VL CDR3s listed in Table2 or Table 3.

[0147] In another embodiment, a nucleic acid molecule(s) of theinvention encodes an antibody or fragment thereof comprising a VH domainhaving an amino acid sequence of any one of the VH domains listed inTable 2 and a VL domain having an amino acid sequence of any one of theVL domains listed in Table 2. In another embodiment, a nucleic acidmolecule(s) of the invention encodes an antibody or fragment thereofcomprising a VH CDR1, a VL CDR1, a VH CDR2, a VL CDR2, a VH CDR3, a VLCDR3, or any combination thereof having an amino acid sequence listed inTable 2. In another embodiment, a nucleic acid molecule(s) of theinvention encodes an antibody or fragment thereof comprising a VH CDR1,a VL CDR1, a VH CDR2, a VL CDR2, a VH CDR3, a VL CDR3, or anycombination thereof having an amino acid sequence listed in Table 3. Inanother embodiment, a nucleic acid molecule(s) of the invention encodesan antibody or fragment thereof comprising a VH CDR1, a VL CDR1, a VHCDR2, a VL CDR2, a VH CDR3, a VL CDR3, or any combination thereof havingan amino acid sequence listed in Table 2 and Table 3.

[0148] The present invention also provides antibodies or fragmentsthereof comprising derivatives of the VH domains, VH CDRs, VL domains,and VL CDRs described herein that immunospecifically bind to an RSVantigen. The present invention also provides antibodies or fragmentsthereof comprising derivatives of SYNAGIS(R), AFFF, Pf12, P12f4, P11d4,A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6,DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S, wherein said antibodies or fragmentsthereof immunospecifically bind to one or more RSV antigens. Standardtechniques known to those of skill in the art can be used to introducemutations in the nucleotide sequence encoding a molecule of theinvention, including, for example, site-directed mutagenesis andPCR-mediated mutagenesis which results in amino acid substitutions.Preferably, the derivatives include less than 25 amino acidsubstitutions, less than 20 amino acid substitutions, less than 15 aminoacid substitutions, less than 10 amino acid substitutions, less than 5amino acid substitutions, less than 4 amino acid substitutions, lessthan 3 amino acid substitutions, or less than 2 amino acid substitutionsrelative to the original molecule. In a preferred embodiment, thederivatives have conservative amino acid substitutions are made at oneor more predicted non-essential amino acid residues. A “conservativeamino acid substitution” is one in which the amino acid residue isreplaced with an amino acid residue having a side chain with a similarcharge. Families of amino acid residues having side chains with similarcharges have been defined in the art. These families include amino acidswith basic side chains (e.g., lysine, arginine, histidine), acidic sidechains (e.g., aspartic acid, glutamic acid), uncharged polar side chains(e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine,cysteine), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine, tryptophan),beta-branched side chains (e.g., threonine, valine, isoleucine) andaromatic side chains (e.g., tyrosine, phenylalanine, tryptophan,histidine). Alternatively, mutations can be introduced randomly alongall or part of the coding sequence, such as by saturation mutagenesis,and the resultant mutants can be screened for biological activity toidentify mutants that retain activity. Following mutagenesis, theencoded protein can be expressed and the activity of the protein can bedetermined.

[0149] In a specific embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises a nucleotidesequence that hybridizes to the nucleotide sequence(s) encodingSYNAGIS®, AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4,A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, or A4B4-F52S understringent conditions, e.g., hybridization to filter-bound DNA in6×sodium chloride/sodium citrate (SSC) at about 45° C. followed by oneor more washes in 0.2×SSC/0.1% SDS at about 50-65° C., under highlystringent conditions, e.g., hybridization to filter-bound nucleic acidin 6×SSC at about 45° C. followed by one or more washes in 0.1×SSC/0.2%SDS at about 68° C., or under other stringent hybridization conditionswhich are known to those of skill in the art (see, for example, Ausubel,F. M. et al., eds., 1989, Current Protocols in Molecular Biology, Vol.1, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., NewYork at pages 6.3.1-6.3.6 and 2.10.3).

[0150] In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence that is at least 35%, at least 40%, at least 45%, at least 50%,at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, or at least 99%identical to the amino acid sequence of SYNAGIS®, AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9,Y1OH6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S.

[0151] In a specific embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence of a VH domain or an amino acid sequence a VL domain encoded bya nucleotide sequence that hybridizes to the nucleotide sequenceencoding any one of the VH or VL domains listed in Table 2 understringent conditions, e.g., hybridization to filter-bound DNA in 6xsodium chloride/sodium citrate (SSC) at about 45° C. followed by one ormore washes in 0.2×SSC/0.1% SDS at about 50-65° C., under highlystringent conditions, e.g., hybridization to filter-bound nucleic acidin 6×SSC at about 45° C. followed by one or more washes in 0.1×SSC/0.2%SDS at about 68° C., or under other stringent hybridization conditionswhich are known to those of skill in the art (see, for example, Ausubel,F. M. et al., eds. , 1989, Current Protocols in Molecular Biology, Vol.1, Green Publishing Associates, Inc. and John Wiley & Sons, Inc., NewYork at pages 6.3.1-6.3.6 and 2.10.3). In another embodiment, anantibody or fragment thereof that immunospecifically binds to a RSVantigen comprises an amino acid sequence of a VH CDR or an amino acidsequence of a VL CDRs encoded by a nucleotide sequence that hybridizesto the nucleotide sequence encoding any one of the VH CDRs or VL CDRslisted in Table 2 or Table 3 under stringent conditions e.g.,hybridization to filter-bound DNA in 6×sodium chloride/sodium citrate(SSC) at about 45° C. followed by one or more washes in 0.2×SSC/0.1% SDSat about 50-65° C., under highly stringent conditions, e.g.,hybridization to filter-bound nucleic acid in 6×SSC at about 45° C.followed by one or more washes in 0.1×SSC/0.2% SDS at about 68° C., orunder other stringent hybridization conditions which are known to thoseof skill in the art. In yet another embodiment, an antibody or fragmentthereof that immunospecifically binds to a RSV antigen comprises anamino acid sequence of a VH CDR and an amino acid sequence of a VL CDRencoded by nucleotide sequences that hybridizes to the nucleotidesequences encoding any one of the VH CDRs and VL CDRs, respectively,listed in Table 2 or Table 3 under stringent conditions, e.g.,hybridization to filter-bound DNA in 6×sodium chloride/sodium citrate(SSC) at about 45° C. followed by one or more washes in 0.2×SSC/0.1% SDSat about 50-65° C., under highly stringent conditions, e.g.,hybridization to filter-bound nucleic acid in 6×SSC at about 45° C.followed by one or more washes in 0.1×SSC/0.2% SDS at about 68° C., orunder other stringent hybridization conditions which are known to thoseof skill in the art .

[0152] In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence of a VH domain that is at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, or at least 99% identical to any one of the VH domains listed inTable 2. In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence of one or more VH CDRs that are at least 35%, at least 40%, atleast 45%, at least 50%, at least 55%, at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, or at least 99% identical to any of the VH CDRs listed inTable 2 or Table 3.

[0153] In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence of a VL domain that is at least 35%, at least 40%, at least45%, at least 50%, at least 55%, at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, or at least 99% identical to any one of the VL domains listed inTable 2. In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence of one or more VL CDRs that are at least 35%, at least 40%, atleast 45%, at least 50%, at least 55%, at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, or at least 99% identical to any of the VL CDRs listed inTable 2 or Table 3.

[0154] In another embodiment, an antibody or fragment thereof thatimmunospecifically binds to a RSV antigen comprises an amino acidsequence that is at least 35%, at least 40%, at least 45%, at least 50%,at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, or at least 99%identical to an amino acid sequence of SYNAGIS®, AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9,Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S

[0155] The present invention also encompasses antibodies or fragmentsthereof that compete with an antibody or Fab fragment listed in Table 2for binding to a RSV antigen. In particular, the present inventionencompasses antibodies or fragments thereof that compete with SYNAGIS®or an antigen-binding fragment thereof for binding to the RSV Fglycoprotein. The present invention also encompasses VL domains, VHdomains, VL CDRs, and VH CDRs that compete with a VL domain, VH domain,VL CDR, or VH CDR listed in Table 2 for binding to a RSV antigen.Further, the present invention encompasses VL CDRs and VL CDRs thatcompete with a VL CDR or VH CDR listed in Table 3 for binding to a RSVantigen.

[0156] The antibodies of the invention include derivatives that aremodified, i. e, by the covalent attachment of any type of molecule tothe antibody such that covalent attachment. For example, but not by wayof limitation, the antibody derivatives include antibodies that havebeen modified, e.g, by glycosylation, acetylation, pegylation,phosphorylation, amidation, derivatization by known protecting/blockinggroups, proteolytic cleavage, linkage to a cellular ligand or otherprotein, etc. Any of numerous chemical modifications may be carried outby known techniques, including, but not limited to specific chemicalcleavage, acetylation, formylation, metabolic synthesis of tunicamycin,etc. Additionally, the derivative may contain one or more non-classicalamino acids.

[0157] The present invention also provides antibodies of the inventionor fragments thereof that comprise a framework region known to those ofskill in the art. Preferably, the framework region of an antibody of theinvention or fragment thereof is human. In a specific embodiment, anantibody of the invention or fragment thereof comprises the frameworkregion of SYNAGIS®.

[0158] The present invention also provides antibodies of the inventionor fragments thereof that comprise constant regions known to those ofskill in the art. Preferably, the constant regions of an antibody of theinvention or fragment thereof are human. In a specific embodiment, anantibody of the invention or fragment thereof comprises the constantregions of SYNAGIS®.

[0159] The present invention also provides for antibodies or fragmentsthereof that have half-lives in a mammal, preferably a human, of greaterthan 15 days, preferably greater than 20 days, greater than 25 days,greater than 30 days, greater than 35 days, greater than 40 days,greater than 45 days, greater than 2 months, greater than 3 months,greater than 4 months, or greater than 5 months. The increasedhalf-lives of the antibodies of the present invention or fragmentsthereof in a mammal, preferably a human, results in a higher serum titerof said antibodies or antibody fragments in the mammal, and thus,reduces the frequency of the administration of said antibodies orantibody fragments and/or reduces the concentration of said antibodiesor antibody fragments to be administered. Antibodies or fragmentsthereof having increased in vivo half-lives can be generated bytechniques known to those of skill in the art. For example, antibodiesor fragments thereof with increased in vivo half-lives can be generatedby modifying (e.g., substituting, deleting or adding) amino acidresidues identified as involved in the interaction between the Fc domainand the FcRn receptor (see, e.g., PCT Publication No. WO 97/34631, whichis incorporated herein by reference in its entirety). Such antibodies orfragments thereof can be tested for binding activity to RSV antigens aswell as for in vivo efficacy using methods known to those skilled in theart, for example, by immunoassays described herein.

[0160] Further, antibodies or fragments thereof with increased in vivohalf-lives can be generated by attaching to said antibodies or antibodyfragments polymer molecules such as high molecular weightpolyethyleneglycol (PEG). PEG can be attached to said antibodies orantibody fragments with or without a multifunctional linker eitherthrough site-specific conjugation of the PEG to the N- or C- terminus ofsaid antibodies or antibody fragments or via epsilon-amino groupspresent on lysine residues. Linear or branched polymer derivatizationthat results in minimal loss of biological activity will be used. Thedegree of conjugation will be closely monitored by SDS-PAGE and massspectrometry to ensure proper conjugation of PEG molecules to theantibodies. Unreacted PEG can be separated from antibody-PEG conjugatesby, e.g., size exclusion or ion-exchange chromatography.PEG-derivatizated antibodies or fragments thereof can be tested forbinding activity to RSV antigens as well as for in vivo efficacy usingmethods known to those skilled in the art, for example, by immunoassaysdescribed herein.

[0161] The present invention also encompasses antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens, saidantibodies or antibody fragments comprising the amino acid sequence ofSYNAGIS® with mutations (e.g., one or more amino acid substitutions) inthe framework regions. In certain embodiments, antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigenscomprise the amino acid sequence of SYNAGIS® with one or more amino acidresidue substitutions in the framework regions of the VH and/or VLdomains depicted in FIG. 1. In a specific embodiment, antibodies orfragments thereof which immunospecifically bind to one or more RSVantigens comprise the framework regions depicted in FIG. 2.

[0162] The present invention also encompasses antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens, saidantibodies or fragments thereof comprising the amino acid sequence ofSYNAGIS® with mutations (e.g., one or more amino acid residuesubstitutions) in the variable and framework regions.

[0163] The present invention also provides for fusion proteinscomprising an antibody or fragment thereof that immunospecifically bindsto a RSV antigen and a heterologous polypeptide. Preferably, theheterologous polypeptide that the antibody or antibody fragment is fusedto is useful for targeting the antibody to respiratory epithelial cells.

[0164] The present invention also provides for panels of antibodies orfragments thereof that immunospecifically bind to an RSV antigen. Inspecific embodiments, the invention provides for panels of antibodies orfragments thereof having different affinities for an RSV antigen,different specificities for an RSV antigen, or different dissociationrates. The invention provides panels of at least 10, preferably at least25, at least 50, at least 75, at least 100, at least 125, at least 150,at least 175, at least 200, at least 250, at least 300, at least 350, atleast 400, at least 450, at least 500, at least 550, at least 600, atleast 650, at least 700, at least 750, at least 800, at least 850, atleast 900, at least 950, or at least 1000 antibodies or fragmentsthereof. Panels of antibodies can be used, for example, in 96 wellplates for assays such as ELISAs.

[0165] The present invention further provides for compositionscomprising one or more antibodies of the invention or fragments thereof.In a specific embodiment, a composition of the present inventioncomprises SYNAGIS®, AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4,A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, and/or A4B4-F52S. Inanother specific embodiment, a composition of the present inventioncomprises an antigen-binding fragment of SYNAGIS®, AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9,Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S.

[0166] In another embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VH domains having an amino acid sequence of any one of the VHdomains listed in Table 2. In another embodiment, a composition of thepresent invention comprises one or more antibodies or fragments thereofcomprising one or more VH CDR1 s having an amino acid sequence of anyone of the VH CDR1s listed in Table 2 or Table 3. In another embodiment,a composition of the present invention comprises one or more antibodiesor fragments thereof comprising one or more VH CDR2s having an aminoacid sequence of any one of the VH CDR2s listed in Table 2 or Table 3.In a preferred embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VH CDR3s having an amino acid sequence of any one of the VH CDR3slisted in Table 2 or Table 3.

[0167] In another embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VL domains having an amino acid sequence of any one of the VLdomains listed in Table 2. In another embodiment, a composition of thepresent invention comprises one or more antibodies or fragments thereofcomprising one or more VL CDR1s having an amino acid sequence of any oneof the VH CDR1s listed in Table 2 or Table 3. In another embodiment, acomposition of the present invention comprises one or more antibodies orfragments thereof comprising one or more VL CDR2s having an amino acidsequence of any one of the VL CDR2s listed in Table 2 or Table 3. In apreferred embodiment, a composition of the present invention comprisesone or more antibodies or fragments thereof comprising one or more VLCDR3s having an amino acid sequence of any one of the VL CDR3s listed inTable 2 or Table 3.

[0168] In another embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VH domains having an amino acid sequence of any one of the VHdomains listed in Table 2 and one or more VL domains having an aminoacid sequence of any one of the VL domains listed in Table 2. In anotherembodiment, a composition of the present invention comprises one or moreantibodies or fragments thereof comprising one or more VH CDR1s havingan amino acid sequence of any one of the VH CDR1 s listed in Table 2 orTable 3 and one or more VL CDR1 s having an amino acid sequence of anyone of the VL CDR1s listed in Table 2 or Table 3. In another embodiment,a composition of the present invention comprises one or more antibodiesor fragments thereof comprising one or more VH CDR1 s having an aminoacid sequence of any one of the VH CDR1 s listed in Table 2 or Table 3and one or more VL CDR2s having an amino acid sequence of any one of theVL CDR2s listed in Table 2 or Table 3. In another embodiment, acomposition of the present invention comprises one or more antibodies orfragments thereof comprising one or more VH CDR1s having an amino acidsequence of any one of the VH CDR1s listed in Table 2 or Table 3 and oneor more VL CDR3s having an amino acid sequence of any one of the VLCDR3s listed in Table 2 or Table 3.

[0169] In another embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VH CDR2s having an amino acid sequence of any one of the VH CDR2slisted in Table 2 or Table 3 and one or more VL CDR1 s having an aminoacid sequence of any one of the VL CDR1 s listed in Table 2 or Table 3.In another embodiment, a composition of the present invention comprisesone or more antibodies or fragments thereof comprising one or more VHCDR2s having an amino acid sequence of any one of the VH CDR2s listed inTable 2 or Table 3 and one or more VL CDR2s having an amino acidsequence of any one of the VL CDR2s listed in Table 2 or Table 3. Inanother embodiment, a composition of the present invention comprises oneor more antibodies or fragments thereof comprising one or more VH CDR2shaving an amino acid sequence of any one of the VH CDR2s listed in Table2 or Table 3 and one or more VL CDR3s having an amino acid sequence ofany one of the VL CDR3s listed in Table 2 or Table 3.

[0170] In another embodiment, a composition of the present inventioncomprises one or more antibodies or fragments thereof comprising one ormore VH CDR3s having an amino acid sequence of any one of the VH CDR3slisted in Table 2 or Table 3 and one or more VL CDR1 s having an aminoacid sequence of any one of the VL CDR1 s listed in Table 2 or Table 3.In another embodiment, a composition of the present invention comprisesone or more antibodies or fragments thereof comprising one or more VHCDR3s having an amino acid sequence of any one of the VH CDR3s listed inTable 2 or Table 3 and one or more VL CDR2s having an amino acidsequence of any one of the VL CDR2s listed in Table 2 or Table 3. In apreferred embodiment, a composition of the present invention comprisesone or more antibodies or fragments thereof comprising one or more VHCDR3s having an amino acid sequence of any one of the VH CDR3s listed inTable 2 or Table 3 and one or more VL CDR3s having an amino acidsequence of any one of the VL CDR3s listed in Table 2 or Table 3. In yetanother embodiment, a composition of the present invention comprises oneor more fusion proteins of the invention.

[0171] As discussed in more detail below, a composition of the inventionmay be used either alone or in combination with other compositions. Theantibodies or fragments thereof may further be recombinantly fused to aheterologous polypeptide at the N- or C- terminus or chemicallyconjugated (including covalently and non-covalently conjugations) topolypeptides or other compositions. For example, antibodies of thepresent invention may be recombinantly fused or conjugated to moleculesuseful as labels in detection assays and effector molecules such asheterologous polypeptides, drugs, radionuclides, or toxins. See, e.g.,PCT publications WO 92/08495; WO 91/14438; WO 89/12624; U.S. Pat. No.5,314,995; and EP 396,387.

[0172] Antibodies of the present invention or fragments thereof may beused, for example, to purify, detect, and target RSV antigens, in bothin vitro and in vivo diagnostic and therapeutic methods. For example,the antibodies or fragments have use in immunoassays for qualitativelyand quantitatively measuring levels of the RSV in biological samplessuch as sputum. See, e.g., Harlow et al., Antibodies: A LaboratoryManual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988)(incorporated by reference herein in its entirety).

[0173] 5.1.1. Antibody Conjugates

[0174] The present invention encompasses antibodies or fragments thereofrecombinantly fused or chemically conjugated (including both covalentlyand non-covalently conjugations) to a heterologous polypeptide (orportion thereof, preferably at least 10, at least 20, at least 30, atleast 40, at least 50, at least 60, at least 70, at least 80, at least90 or at least 100 amino acids of the polypeptide) to generate fusionproteins. The fusion does not necessarily need to be direct, but mayoccur through linker sequences. For example, antibodies may be used totarget heterologous polypeptides to particular cell types (e.g.,respiratory epithelial cells), either in vitro or in vivo, by fusing orconjugating the antibodies to antibodies specific for particular cellsurface receptors. Antibodies fused or conjugated to heterologouspolypeptides may also be used in in vitro immunoassays and purificationmethods using methods known in the art. See e.g., PCT publication WO93/21232; EP 439,095; Naramura et al., Immunol. Lett. 39:91-99 (1994);U.S. Pat. No. 5,474,981; Gillies et al., PNAS 89:1428-1432 (1992); andFell et al., J. Immunol. 146:2446-2452(1991), which are incorporated byreference in their entireties.

[0175] In one embodiment, a fusion protein of the invention comprisesSYNAGIS®, AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4,A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, or A4B4-F52S and aheterologous polypeptide. In another embodiment, a fusion protein of theinvention comprises an antigen-binding fragment of SYNAGIS®, AFFF, Pf12,P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4,M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S and a heterologous polypeptide. In anotherembodiment, a fusion protein of the invention comprises one or more VHdomains having the amino acid sequence of any one of the VH domainslisted in Table 2 or one or more VL domains having the amino acidsequence of any one of the VL domains listed in Table 2 and aheterologous polypeptide. In another embodiment, a fusion protein of thepresent invention comprises one or more VH CDRs having the amino acidsequence of any one of the VH CDRs listed in Table 2 or Table 3 and aheterologous polypeptide. In another embodiment, a fusion proteincomprises one or more VL CDRs having the amino acid sequence of any oneof the VL CDRs listed in Table 2 or Table 3 and a heterologouspolypeptide. In another embodiment, a fusion protein of the inventioncomprises at least one VH domain and at least one VL domain listed inTable 2 and a heterologous polypeptide. In yet another embodiment, afusion protein of the invention comprises at least one VH CDR and atleast one VL CDR domain listed in Table 2 or Table 3 and a heterologouspolypeptide.

[0176] The present invention further includes compositions comprisingheterologous polypeptides fused or conjugated to antibody fragments. Forexample, the heterologous polypeptides may be fused or conjugated to aFab fragment, Fd fragment, Fv fragment, F(ab)₂ fragment, or portionthereof. Methods for fusing or conjugating polypeptides to antibodyportions are known in the art. See, e.g., U.S. Pat. Nos. 5,336,603,5,622,929, 5,359,046, 5,349,053, 5,447,851, and 5,112,946; EP 307,434;EP 367,166; PCT publication Nos. WO 96/04388 and WO 91/06570; Ashkenaziet al., Proc. Natl. Acad. Sci. USA 88:10535-10539 (1991); Zheng et al.,J. Immunol. 154:5590-5600 (1995); and Vil et al., Proc. Natl. Acad. Sci.USA 89:11337- 11341(1992) (said references incorporated by reference intheir entireties).

[0177] Additional fusion proteins of the invention may be generatedthrough the techniques of gene-shuffling, motif-shuffling,exon-shuffling, and/or codon-shuffling (collectively referred to as “DNAshuffling”). DNA shuffling may be employed to alter the activities ofantibodies of the invention or fragments thereof (e.g., antibodies orfragments thereof with higher affinities and lower dissociation rates).See, generally, U.S. Pat. Nos. 5,605,793; 5,811,238; 5,830,721;5,834,252; and 5,837,458, and Patten et al., Curr. Opinion Biotechnol.8:724-33 (1997); Harayama, Trends Biotechnol. 16(2):76-82 (1998);Hansson, et al., J. Mol. Biol. 287:265-76 (1999); and Lorenzo andBlasco, Biotechniques 24(2):308-13 (1998) (each of these patents andpublications are hereby incorporated by reference in its entirety). Inone embodiment, antibodies or fragments thereof, or the encodedantibodies or fragments thereof, may be altered by being subjected torandom mutagenesis by error-prone PCR, random nucleotide insertion orother methods prior to recombination. In another embodiment, one or moreportions of a polynucleotide encoding an antibody or antibody fragment,which portions immunospecifically bind to a RSV antigen may berecombined with one or more components, motifs, sections, parts,domains, fragments, etc. of one or more heterologous molecules.

[0178] Moreover, the antibodies of the present invention or fragmentsthereof can be fused to marker sequences, such as a peptide tofacilitate purification. In preferred embodiments, the marker amino acidsequence is a hexa-histidine peptide, such as the tag provided in a pQEvector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, Calif., 91311),among others, many of which are commercially available. As described inGentz et al., 1989, Proc. Natl. Acad. Sci. USA 86:821-824, for instance,hexa-histidine provides for convenient purification of the fusionprotein. Other peptide tags useful for purification include, but are notlimited to, the hemagglutinin“HA” tag, which corresponds to an epitopederived from the influenza hemagglutinin protein (Wilson et al., 1984,Cell 37:767) and the “flag” tag.

[0179] The present invention further encompasses antibodies or fragmentsthereof conjugated to a diagnostic or therapeutic agent. The antibodiescan be used diagnostically to, for example, monitor the development orprogression of a RSV infection as part of a clinical testing procedureto, e.g., determine the efficacy of a given treatment regimen. Detectioncan be facilitated by coupling the antibody or fragment thereof to adetectable substance. Examples of detectable substances include variousenzymes, prosthetic groups, fluorescent materials, luminescentmaterials, bioluminescent materials, radioactive materials, positronemitting metals, and nonradioactive paramagnetic metal ions. Thedetectable substance may be coupled or conjugated either directly to theantibody (or fragment thereof) or indirectly, through an intermediate(such as, for example, a linker known in the art) using techniques knownin the art. See, for example, U.S. Pat. No. 4,741,900 for metal ionswhich can be conjugated to antibodies for use as diagnostics accordingto the present invention. Examples of suitable enzymes includehorseradish peroxidase, alkaline phosphatase, beta-galactosidase, oracetylcholinesterase; examples of suitable prosthetic group complexesinclude streptavidin/biotin and avidin/biotin; examples of suitablefluorescent materials include umbelliferone, fluorescein, fluoresceinisothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansylchloride or phycoerythrin; an example of a luminescent material includesluminol; examples of bioluminescent materials include luciferase,luciferin, and aequorin; and examples of suitable radioactive materialinclude ¹²⁵I, ^(1311,) ¹¹¹In or ⁹⁹Tc.

[0180] An antibody or fragment thereof may be conjugated to atherapeutic moiety such as a cytotoxin, e.g., a cytostatic or cytocidalagent, a therapeutic agent or a radioactive metal ion, e.g.,alpha-emitters. A cytotoxin or cytotoxic agent includes any agent thatis detrimental to cells. Examples include paclitaxol, cytochalasin B,gramicidin D, ethidium bromide, emetine, mitomycin, etoposide,tenoposide, vincristine, vinblastine, colchicin, doxorubicin,daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin,actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine,tetracaine, lidocaine, propranolol, and puromycin and analogs orhomologs thereof. Therapeutic agents include, but are not limited to,antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine,cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g.,mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) andlomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol,streptozotocin, mitomycin C, and cisdichlorodiamine platinum (II) (DDP)cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) anddoxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin),bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents(e.g., vincristine and vinblastine).

[0181] Further, an antibody or fragment thereof may be conjugated to atherapeutic agent or drug moiety that modifies a given biologicalresponse. Therapeutic agents or drug moieties are not to be construed aslimited to classical chemical therapeutic agents. For example, the drugmoiety may be a protein or polypeptide possessing a desired biologicalactivity. Such proteins may include, for example, a toxin such as abrin,ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such astumor necrosis factor, α-interferon, β-interferon, nerve growth factor,platelet derived growth factor, tissue plasminogen activator, anapoptotic agent, e.g., TNF-α, TNF-β, AIM I (see, InternationalPublication No. WO 97/33899), AIM II (see, International Publication No.WO 97/34911), Fas Ligand (Takahashi et al., 1994, J. Iminunol.,6:1567-1574), and VEGI (see, International Publication No. WO 99/23105),a thrombotic agent or an anti-angiogenic agent, e.g., angiostatin orendostatin; or, a biological response modifier such as, for example, alymphokine (e.g., interleukin-1 (“IL-1”), interleukin-2 (“IL-2”),interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor(“GM-CSF”), and granulocyte colony stimulating factor (“G-CSF”)), or agrowth factor (e.g., growth hormone (“GH”)).

[0182] Techniques for conjugating such therapeutic moieties toantibodies are well known, see, e.g., Arnon et al., “MonoclonalAntibodies For Immunotargeting Of Drugs In Cancer Therapy”, inMonoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp.243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For DrugDelivery”, in Controlled Drug Delivery (2nd Ed.), Robinson et al.(eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “AntibodyCarriers Of Cytotoxic Agents In Cancer Therapy: A Review”, in MonoclonalAntibodies '84: Biological And Clinical Applications, Pinchera et al.(eds.), pp. 475-506 (1985); “Analysis, Results, And Future ProspectiveOf The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, inMonoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al.(eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., 1982,Immunol. Rev. 62:119-58.

[0183] An antibody or fragment thereof, with or without a therapeuticmoiety conjugated to it, administered alone or in combination withcytotoxic factor(s) and/or cytokine(s) can be used as a therapeutic.

[0184] Alternatively, an antibody can be conjugated to a second antibodyto form an antibody heteroconjugate as described by Segal in U.S. PatentNo. 4,676,980, which is incorporated herein by reference in itsentirety.

[0185] Antibodies may also be attached to solid supports, which areparticularly useful for immunoassays or purification of the targetantigen. Such solid supports include, but are not limited to, glass,cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride orpolypropylene.

[0186] 5.2. Prophylactic and Therapeutic Uses of Antibodies

[0187] The present invention is directed to antibody-based therapieswhich involve administering antibodies of the invention or fragmentsthereof to a mammal, preferably a human, for preventing, treating, orameliorating one or more symptoms associated with a RSV infection.Prophylactic and therapeutic compounds of the invention include, but arenot limited to, antibodies of the invention (including fragments,analogs and derivatives thereof as described herein) and nucleic acidsencoding antibodies of the invention (including fragments, analogs andderivatives thereof and anti-idiotypic antibodies as described herein).Antibodies of the invention or fragments thereof may be provided inpharmaceutically acceptable compositions as known in the art or asdescribed herein.

[0188] Antibodies of the present invention or fragments thereof thatfunction as antagonists of a RSV infection can be administered to amammal, preferably a human, to treat, prevent or ameliorate one or moresymptoms associated with a RSV infection. For example, antibodies orfragments thereof which disrupt or prevent the interaction between a RSVantigen and its host cell receptor may be administered to a mammal,preferably a human, to treat, prevent or ameliorate one or more symptomsassociated with a RSV infection.

[0189] In a specific embodiment, an antibody or fragment thereofprevents RSV from binding to its host cell receptor by at least 99%, atleast 95%, at least 90%, at least 85%, at least 80%, at least 75%, atleast 70%, at least 60%, at least 50%, at least 45%, at least 40%, atleast 45%, at least 35%, at least 30%, at least 25%, at least 20%, or atleast 10% relative to RSV binding to its host cell receptor in theabsence of said antibodies or antibody fragments. In another embodiment,a combination of antibodies, a combination of antibody fragments, or acombination of antibodies and antibody fragments prevent RSV frombinding to its host cell receptor by at least 99%, at least 95%, atleast 90%, at least 85%, at least 80%, at least 75%, at least 70%, atleast 60%, at least 50%, at least 45%, at least 40%, at least 45%, atleast 35%, at least 30%, at least 25%, at least 20%, or at least 10%relative to RSV binding to its host cell receptor in the absence of saidantibodies and/or antibody fragments.

[0190] Antibodies or fragments thereof which do not prevent RSV frombinding its host cell receptor but inhibit or downregulate RSVreplication can also be administered to a mammal to treat, prevent orameliorate one or more symptoms associated with a RSV infection. Theability of an antibody or fragment thereof to inhibit or downregulateRSV replication may be determined by techniques described herein orotherwise known in the art. For example, the inhibition ordownregulation of RSV replication can be determined by detecting the RSVtiter in the lungs of a mammal, preferably a human.

[0191] In a specific embodiment, an antibody of the present invention orfragment thereof inhibits or downregulates RSV replication by at least99%, at least 95%, at least 90%, at least 85%, at least 80%, at least75%, at least 70%, at least 60%, at least 50%, at least 45%, at least40%, at least 45%, at least 35%, at least 30%, at least 25%, at least20%, or at least 10% relative to RSV replication in absence of saidantibodies or antibody fragments. In another embodiment, a combinationof antibodies, a combination of antibody fragments, or a combination ofantibodies and antibody fragments inhibit or downregulate RSVreplication by at least 99%, at least 95%, at least 90%, at least 85%,at least 80%, at least 75%, at least 70%, at least 60%, at least 50%, atleast 45%, at least 40%, at least 45%, at least 35%, at least 30%, atleast 25%, at least 20%, or at least 10% relative to RSV replication inabsence of said antibodies and/or antibody fragments.

[0192] One or more antibodies of the present invention or fragmentsthereof that immunospecifically bind to one or more RSV antigens may beused locally or systemically in the body as a therapeutic. Theantibodies of this invention or fragments thereof may also beadvantageously utilized in combination with other monoclonal or chimericantibodies, or with lymphokines or hematopoietic growth factors (suchas, e.g., IL-2, IL-3 and IL-7), which, for example, serve to increasethe number or activity of effector cells which interact with theantibodies. The antibodies of this invention or fragments thereof mayalso be advantageously utilized in combination with other monoclonal orchimeric antibodies, or with lymphokines or hematopoietic growth factors(such as, e.g., IL-2, IL-3 and IL-7), which, for example, serve toincrease the immune response. The antibodies of this invention orfragments thereof may also be advantageously utilized in combinationwith one or more drugs used to treat RSV infection such as, for exampleanti-viral agents. Antibodies of the invention or fragments may be usedin combination with one or more of the following drugs: NIH-351 (GeminiTechnologies), recombinant RSV vaccine (Aviron), RSVf-2 (Intracel),F-50042 (Pierre Fabre), T-786 (Trimeris), VP-36676 (ViroPharma), RFI-641(American Home Products), VP-14637 (ViroPharma), PFP-1 and PFP-2(American Home Products), RSV vaccine (Avant Immunotherapeutics), andF-50077 (Pierre Fabre).

[0193] The antibodies of the invention may be administered alone or incombination with other types of treatments (e.g., hormonal therapy,immunotherapy, and anti-inflammatory agents). Generally, administrationof products of a species origin or species reactivity (in the case ofantibodies) that is the same species as that of the patient ispreferred. Thus, in a preferred embodiment, human or humanizedantibodies, fragments derivatives, analogs, or nucleic acids, areadministered to a human patient for therapy or prophylaxis.

[0194] It is preferred to use high affinity and/or potent in vivoinhibiting antibodies and/or neutralizing antibodies thatimmunospecifically bind to a RSV antigen, for both immunoassays directedto RSV, prevention of RSV infection and therapy for RSV infection. It isalso preferred to use polynucleotides encoding high affinity and/orpotent in vivo inhibiting antibodies and/or neutralizing antibodies thatimmunospecifically bind to a RSV antigen, for both immunoassays directedto RSV and therapy for RSV infection. Such antibodies or fragmentsthereof will preferably have an affinity for the RSV F glycoproteinand/or fragments of the F glycoprotein.

[0195] In one embodiment, therapeutic or pharmaceutical compositionscomprising antibodies of the invention or fragments thereof areadministered to a mammal, preferably a human, to treat, prevent orameliorate one or more symptoms associated with RSV infection. Inanother embodiment, therapeutic or pharmaceutical compositionscomprising antibodies of the invention or fragments thereof areadministered to a human with cystic fibrosis, bronchopulmonarydysplasia, congenital heart disease, congenital immunodeficiency oracquired immunodeficiency, or to a human who has had a bone marrowtransplant to treat, prevent or ameliorate one or more symptomsassociated with RSV infection. In another embodiment, therapeutic orpharmaceutical compositions comprising antibodies of the invention orfragments thereof are administered to a human infant, preferably a humaninfant born prematurely or a human infant at risk of hospitalization forRSV infection to treat, prevent or ameliorate one or more symptomsassociated with RSV infection. In yet another embodiment, therapeutic orpharmaceutical compositions comprising antibodies of the invention orfragments thereof are administered to the elderly or people in grouphomes (e.g., nursing homes or rehabilitation centers).

[0196] In a specific embodiment, a mammal, preferably a human, isadministered a therapeutic or pharmaceutical composition comprising oneor more antibodies of the present invention or fragments thereof for thetreatment, prevention or amelioration of one or more symptoms associatedwith a RSV infection in an amount effective for decreasing RSV titers.In accordance with this embodiment, an effective amount of antibodies orantibody fragments reduces the RSV titers in the lung as measured, forexample, by the concentration of RSV in sputum samples or a lavage fromthe lungs from a mammal. In another embodiment, a mammal, preferably ahuman, is administered a therapeutic or pharmaceutical compositioncomprising one or more antibodies of the present invention or fragmentsthereof for the treatment, prevention or amelioration of symptomsassociated with a RSV infection in an amount effective for inducing animmune response in the mammal.

[0197] In another embodiment, a mammal, preferably a human, isadministered a first dose of a therapeutic or pharmaceutical compositioncomprising less than 15 mg/kg, preferably less than 10 mg/kg, less than5 mg/kg, less than 3 mg/kg, less than 1 mg/kg or less than 0.5 mg/kg ofone or more antibodies or fragments thereof that immunospecifically bindto one or more RSV antigens with higher affinity and/or higher aviditythan previously known antibodies (e.g., SYNAGIS®) for the prevention,treatment or amelioration of one or more symptoms associated with a RSVinfection in an amount effective to induce a serum titer of at least 1μg/ml, preferably at least 2 μg/ml, at least 5 μg/ml, at least 10 μg/ml,at least 15 μg/ml, at least 20 μg/ml, or at least 25 μg/ml 20 days(preferably 25, 30, 35, 40 days) after the administration of the firstdose and prior to the administration of a subsequent dose. Preferably,the serum titer of said antibodies or antibody fragments is less than 30μg/ml 30 days after the administration of the first dose and prior tothe administration of a subsequent dose. Preferably, said antibodies areAFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7,1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10,A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, and A4B4-F52S

[0198] In another embodiment, a mammal, preferably a human, isadministered a first dose of a therapeutic or pharmaceutical compositioncomprising less than 15 mg/kg, preferably less than 5 mg/kg, less than 3mg/kg, less than 1 mg/kg or less than 0.5 mg/kg of one or moreantibodies or fragments thereof which have increased in vivo half-livesand which immunospecifically bind to one or more RSV antigens withhigher affinity and/or higher avidity than previously known antibodies(e.g., SYNAGIS®) for the prevention, treatment or amelioration of one ormore symptoms associated with a RSV infection in an amount effective toinduce a serum titer of at least 1 μg/ml, preferably at least 2 μg/ml,at least 5 μg/ml, at least 10 μg/ml, at least 15 μg/ml, at least 20μg/ml, or at least 25 μg/ml 25 days (preferably 30, 35, or 40 days)after the administration of the first dose and prior to theadministration of a subsequent dose. Preferably, the serum titer of saidantibodies or antibody fragments is less than 30 μg/ml 30 days after theadministration of the first dose and prior to the administration of asubsequent dose. Preferably, the novel antibodies are AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9,Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-1SB10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, and A4B4-F52S.

[0199] In another embodiment, a mammal, preferably a human, isadministered a first dose of a therapeutic or pharmaceutical compositioncomprising approximately 15 mg/kg of HL-SYNAGIS or antigen-bindingfragments thereof for the prevention, treatment or amelioration of oneor more symptoms associated with a RSV infection in an amount effectiveto induce a serum titer of at least 1 μg/ml, preferably at least 30μg/ml, at least 35 μg/ml, at least 40 μg/ml, or at least 50 μg/ml 25days (preferably 30, 35, or 40 days) after the administration of thefirst dose and prior to the administration of a subsequent dose. Inanother embodiment, a mammal, preferably a human, is administered afirst dose of a therapeutic or pharmaceutical composition comprisingless than 15 mg/kg (preferably 10 mg/kg or less, 5 mg/kg or less, 3mg/kg or less, 1 mg/kg or less, or 0.5 mg/kg or less) of HL-SYNAGIS orantigen-binding fragments thereof for the prevention, treatment oramelioration of one or more symptoms associated with a RSV infection inan amount effective to induce a serum titer of at least 1 μg/ml,preferably at least 30 μg/ml, at least 35 μg/ml, at least 40 μg/ml, orat least 50 μg/ml 25 days (preferably 30, 35, or 40 days) after theadministration of the first dose and prior to the administration of asubsequent dose.

[0200] The present invention encompasses therapeutic or pharmaceuticalcompositions for pulmonary delivery comprising one or more antibodies orfragments thereof which immunospecifically bind to one or more RSVantigens with a higher affinity and/or a higher avidity than previouslyknown antibodies (e.g., SYNAGIS®). The present invention alsoencompasses therapeutic or pharmaceutical compositions for pulmonarydelivery comprising SYNAGIS® or an antigen-binding fragment thereof.

[0201] In one embodiment, a mammal, preferably a human, is administereda first dose of a therapeutic or pharmaceutical composition forpulmonary delivery comprising less than 15 mg/kg, preferably less than 5mg/kg, less than 3 mg/kg, less than 1 mg/kg or less than 0.5 mg/kg, orless than 0.01 mg/kg of one or more antibodies or fragments thereofwhich immunospecifically bind to one or more RSV antigens with higheraffinity and/or higher avidity than previously known antibodies (e.g.,SYNAGIS®) for the prevention, treatment or amelioration of one or moresymptoms associated with a RSV infection in an amount effective toinduce a titer of 20 ng per mg of lung protein (preferably at least 40ng/mg, at least 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, at least75 ng/mg, at least 100 ng/mg, or at least 150 ng/mg) in an intubationsample or lavage from the lungs of said mammal 20 days (preferably 25,30, 35, or 40 days) after the administration of the first dose and priorto the administration of a subsequent dose. Preferably, the serum titerof said antibodies or antibody fragments is less than 100 ng/ml ofprotein 30 days after the administration of the first dose and prior tothe administration of a subsequent dose. Preferably, the novelantibodies are AFFF, Pf12, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4,A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5,L2-15B10, A13a11, A1h5, A4B4(1), A4B4L1FR-S28R, and A4B4-F52S.

[0202] In another embodiment, a mammal, preferably a human, isadministered a first dose of a therapeutic or pharmaceutical compositionfor pulmonary delivery comprising approximately 15 mg/kg of SYNAGIS® orfragments thereof for the prevention, treatment or amelioration of oneor more symptoms associated with a RSV infection in an amount effectiveto induce a titer of 20 ng per mg of lung protein (preferably at least40 ng/mg, at least 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, atleast 75 ng/mg, at least 100 ng/mg, or at least 150 ng/mg) an intubationsample or lavage from the lungs of said mammal 30 days (preferably 35 or40 days) after the administration of the first dose and prior to theadministration of a subsequent dose. In another embodiment, a mammal,preferably a human, is administered a first dose of a therapeutic orpharmaceutical composition for pulmonary delivery comprising less than15 mg/kg (preferably 10 mg/kg or less, 5 mg/kg or less, 3 mg/kg or less,1 mg/kg or less, or 0.5 mg/kg or less) of SYNAGIS®or fragments thereoffor the prevention of a RSV infection in an amount effective to induce atiter of 20 ng per mg of lung protein (preferably at least 40 ng/mg, atleast 60 ng/mg, at least 80 ng/mg, at least 50 ng/mg, at least 75 ng/mg,at least 100 ng/mg, or at least 150 ng/mg) in an intubation sample orlavage from the lungs of said mammal 30 days (preferably 35 or 40 days)after the administration of the first dose and prior to theadministration of a subsequent dose.

[0203] The present invention encompasses therapeutic or pharmaceuticalcompositions for pulmonary delivery comprising one or more antibodies orfragments thereof which have increased in vivo half-lives and whichimmunospecifically bind to one or more RSV antigens with a higheraffinity and/or a higher avidity than previously known antibodies (e.g.,SYNAGIS®). The present invention also encompasses therapeutic orpharmaceutical compositions for pulmonary delivery comprising HL-SYNAGISor an antigen-binding fragment thereof.

[0204] The present invention encompasses sustained release compositionscomprising one or more antibodies or fragments thereof which haveincreased in vivo half-lives and which immunospecifically bind to one ormore RSV antigens with a higher affinity and/or a higher avidity thanpreviously known antibodies (e.g., SYNAGIS®). The present invention alsoencompasses sustained release compositions comprising SYNAGIS® or anantigen-binding fragment thereof.

[0205] In one embodiment, a mammal, preferably a human, is administereda first dose of a sustained release formulation comprising less than 15mg/kg, preferably less than 5 mg/kg, less than 3 mg/kg, less than 1mg/kg or less than 0.5 mg/kg of one or more antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens withhigher affinity and/or higher avidity than previously known antibodies(e.g., SYNAGIS®) for the prevention, treatment or amelioration of one ormore symptoms associated with a RSV infection in an amount effective toinduce a serum titer of at least 1 μ/ml, preferably at least 2 μg/ml, atleast 5 μg/ml, at least 10 μg/ml, at least 15 μg/ml, at least 20 μg/ml,or at least 25 μg/ml for at least 10 days (preferably at least 15, atleast 20, at least 25, at least 30, at least 35, or at least 40 days)after the administration of the first dose and prior to theadministration of a subsequent dose. Preferably, the serum titer of saidantibodies or antibody fragments is less than 30 μg/ml 30 days after theadministration of the first dose and prior to the administration of asubsequent dose. Preferably, the novel antibodies are AFFF, Pf12, P12f4,P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9,Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4L1FR-S28R, or A4B4-F52S.

[0206] In another embodiment, a mammal, preferably a human, isadministered a first dose of a sustained release formulation comprisingless than 15 mg/kg, preferably less than 5 mg/kg, less than 3 mg/kg,less than 1 mg/kg or less than 0.5 mg/kg of one or more antibodies orfragments thereof which immunospecifically bind to one or more RSVantigens with higher affinity and/or higher avidity than previouslyknown antibodies (e.g., SYNAGIS®) for the prevention, treatment, oramelioration of one or more symptoms associated with a RSV infection inan amount effective to induce a serum titer of 1 μg/ml, preferably 2μg/ml, 5 μg/ml, 10 μg/ml, 15 μg/ml, 20 μg/ml, or 25 μg/ml that ismaintained for at least 10 days (preferably at least 15, at least 20, atleast 25, at least 30, at least 35, or at least 40 days) after theadministration of the first dose and prior to the administration of asubsequent dose without exceeding a serum titer of 30 μg/ml.

[0207] In another embodiment, a mammal, preferably a human, isadministered a first dose of a sustained release formulation comprisingapproximately 15 mg/kg of SYNAGIS® or fragments thereof for theprevention, treatment or amelioration of one or more symptoms associatedwith a RSV infection in an amount effective to induce a titer of atleast 30 μg/ml, preferably at least 35 μg/ml, at least 40 μg/ml, or atleast 50 μg/ml 25 days (preferably 30, 35, or 40 days) after theadministration of the first dose and prior to the administration of asubsequent dose. In another embodiment, a mammal, preferably a human, isadministered a first dose of a sustained release formulation comprisingless than 15 mg/kg (preferably 10 mg/kg or less, 5 mg/kg or less, 3mg/kg or less, 1 mg/kg or less, or 0.5 mg/kg or less) of SYNAGIS® orfragments thereof for the prevention of a RSV infection in an amounteffective to induce at least 30 μg/ml, preferably at least 35 μg/ml, atleast 40 μg/ml, or at least 50 μg/ml 25 days (preferably 30, 35, or 40days) after the administration of the first dose and prior to theadministration of a subsequent dose.

[0208] In another embodiment, a mammal, preferably a human, isadministered a first dose of a sustained release formulation comprisingless than 15 mg/kg, preferably less than 5 mg/kg, less than 3 mg/kg,less than 1 mg/kg or less than 0.5 mg/kg of SYNAGIS® or anantigen-binding fragment thereof for the prevention, treatment, oramelioration of one or more symptoms associated with a RSV infection inan amount effective to induce a serum titer of 1 μg/ml, preferably 2μg/ml, 5 μg/ml, 10 μg/ml, 15 μg/ml, 20 μg/ml, or 25 μg/ml that ismaintained for at least 10 days (preferably at least 15, at least 20, atleast 25, at least 30, at least 35, or at least 40 days) after theadministration of the first dose and prior to the administration of asubsequent dose without exceeding a serum titer of 30 μg/ml.

[0209] The present invention encompasses sustained release formulationscomprising one or more antibodies or fragments thereof which haveincreased in vivo half-lives and which immunospecifically bind to one ormore RSV antigens with a higher affinity and/or a higher avidity thanpreviously known antibodies such as, e.g, SYNAGIS®. The presentinvention also encompasses sustained release formulations comprisingHL-SYNAGIS or an antigen-binding fragment thereof.

[0210] The present invention encompasses sustained release formulationsfor pulmonary delivery comprising one or more antibodies or fragmentsthereof which immunospecifically bind to one or more RSV antigens with ahigher affinity and/or a higher avidity than previously known antibodies(e.g., SYNAGIS®). The present invention also encompasses sustainedrelease formulations for pulmonary delivery comprising one or moreantibodies or fragments thereof which have increased in vivo half-livesand which immunospecifically bind to one or more RSV antigens with ahigher affinity and/or a higher avidity than previously known antibodies(e.g., SYNAGIS®). The present invention also encompasses sustainedrelease formulations for pulmonary delivery comprising SYNAGIS® orfragments thereof. The present invention further encompasses sustainedrelease formulations for pulmonary delivery comprising HL-SYNAGIS or anantigen-binding fragment thereof.

[0211] In another embodiment, a mammal, preferably a human, isadministered a first dose of a therapeutic or pharmaceutical compositioncomprising less than 10 mg/kg, less than 5 mg/kg, less than 3 mg/kg,less than 1 mg/kg or less than 0.5 mg/kg of one or more antibodies ofthe present invention or fragments thereof for the prevention, treatmentor amelioration of one or more symptoms associated with a RSV infectionin an amount effective to induce a serum titer of at least 35 μg/ml, atleast 40 μg/ml, at least 50 μg/ml, at least 80 μg/ml, at least 100μg/ml, at least 120 μg/ml, at least 150 μg/ml, at least 200 μg/ml, atleast 250 μg/ml, or at least 300 μg/ml 20 days (preferably 25, 30, 35 or40 days) after the administration of the first dose. In anotherembodiment, a mammal, preferably a human, is administered a first doseof a therapeutic or pharmaceutical composition comprising approximately15 mg/kg of one or more antibodies of the present invention or fragmentsthereof for the prevention, treatment or amelioration of one or moresymptoms associated with a RSV infection in an amount effective toinduce a serum titer of at least 100 μg/ml, at least 125 μg/ml, at least150 μg/ml, at least 200 μg/ml, at least 250 μg/ml, at least 300 μg/ml,at least 350 μg/ml, at least 400 μg/ml, or at least 450 μg/ml 20 days(preferably 25, 30, 35 or 40 days) after the administration of the firstdose. The term “approximately 15 mg/kg” as used herein refers to a rangeof between 14 mg/kg and 16 mg/kg.

[0212] In another embodiment, a mammal, preferably a human, isadministered a dose of a pharmaceutical composition comprising one ormore antibodies of the present invention or fragments thereof for theprevention a RSV infection in an amount effective to induce aprophylactically effective serum titer of less than 10 μ/ml, less than 8μg/ml, less than 5 μg/ml, less than 3 μg/ml, less than 1 μg/ml, or lessthan 0.5 μg/ml 30 days after the administration of the dose, whereinsaid prophylactically effective serum titer is the serum titer thatreduces the incidence of RSV infection in the human or the serum titerin a cotton rat that results in a RSV titer 5 days after challenge with10⁵ pfu RSV that is 99% lower than the RSV titer in the cotton rat 5days after challenge with 10⁵ pfu of RSV in a cotton rat notadministered the dose prior to challenge. Preferably, the dose of thepharmaceutical composition comprises less than 10 mg/kg, less than 5mg/kg, less than 3 mg/kg, less than 1 mg/kg, or less than 0.5 mg/kg ofone or more antibodies of the present invention or fragments thereof.

[0213] In yet another embodiment, a mammal, preferably a human, isadministered a dose of a therapeutic or pharmaceutical compositioncomprising one or more antibodies of the present invention or fragmentsthereof for the treatment or amelioration of one or more symptomsassociated with a RSV infection in an amount effective to induce atherapeutically effective serum titer of less than 10 μg/ml, less than 8μg/ml, less than 5 μg/ml, less than 3 μg/ml, less than 1 μg/ml, or lessthan 0.5 μg/ml 30 days after the administration of the dose, whereinsaid therapeutically effective serum titer is the serum titer thatreduces the severity or length of RSV infection or is the serum titer ina cotton rat that results in a RSV titer in the rat 5 days afterchallenge with 10⁵ pfu RSV that is 99% lower than the RSV titer 5 daysafter challenge with 10⁵ pfu of RSV in a cotton rat not administered thedose prior to challenge. Preferably, the dose of the therapeutic orpharmaceutical composition comprises less than 12 mg/kg, less than 10mg/kg, less than 5 mg/kg, less than 3 mg/kg, less than 1 mg/kg, or lessthan 0.5 mg/kg of one or more antibodies of the present invention orfragments thereof.

[0214] 5.3. Methods of Administration of Antibodies

[0215] The invention provides methods of treatment, prophylaxis, andamelioration of one or more symptoms associated with RSV infection byadministrating to a subject of an effective amount of antibody orfragment thereof, or pharmaceutical composition comprising an antibodyof the invention or fragment thereof. In a preferred aspect, an antibodyor fragment thereof is substantially purified (i.e., substantially freefrom substances that limit its effect or produce undesiredside-effects). The subject is preferably a mammal such as non-primate(e.g., cows, pigs, horses, cats, dogs, rats etc.) and a primate (e.g.,monkey such as a cynomolgous monkey and a human). In a preferredembodiment, the subject is a human. In another preferred embodiment, thesubject is a human infant or a human infant born prematurely. In anotherembodiment, the subject is a human with cystic fibrosis,bronchopulmonary dysplasia, congenital heart disease, congenitalimmunodeficiency or acquired immunodeficiency, a human who has had abone marrow transplant, or an elderly human

[0216] Various delivery systems are known and can be used to administeran antibody of the invention or a fragment thereof, e.g., encapsulationin liposomes, microparticles, microcapsules, recombinant cells capableof expressing the antibody or antibody fragment, receptor-mediatedendocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)),construction of a nucleic acid as part of a retroviral or other vector,etc. Methods of administering an antibody or fragment thereof, orpharmaceutical composition include, but are not limited to, parenteraladministration (e.g., intradermal, intramuscular, intraperitoneal,intravenous and subcutaneous), epidural, and mucosal (e.g., intranasaland oral routes). In a specific embodiment, antibodies of the presentinvention or fragments thereof, or pharmaceutical compositions areadministered intramuscularly, intravenously, or subcutaneously. Thecompositions may be administered by any convenient route, for example byinfusion or bolus injection, by absorption through epithelial ormucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa,etc.) and may be administered together with other biologically activeagents. Administration can be systemic or local. In addition, pulmonaryadministration can also be employed, e.g., by use of an inhaler ornebulizer, and formulation with an aerosolizing agent. See, e.g., U.S.Pat. Nos. 6,019,968, 5,985,320, 5,985,309, 5,934,272, 5,874,064,5,855,913, 5,290,540, and 4,880,078; and PCT Publication Nos. WO92/19244, WO 97/32572, WO 97/44013, WO 98/31346, and WO 99/66903, eachof which is incorporated herein by reference their entirety. In apreferred embodiment, an antibody of the invention or fragment thereof,or composition of the invention is administered using Alkermes AIR™pulmonary drug delivery technology (Alkermes, Inc., Cambridge, Mass.).

[0217] The invention provides for any method of administrating lowerdoses of known antibodies or fragments thereof which immunospecificallybind to one or more RSV antigens than previously thought to be effectivefor the prevention, treatment or amelioration of one or more symptomsassociated with a RSV infection. Preferably, lower doses of knownantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens are administered by pulmonary administration. Thepresent invention also provides for any method of administering a novelantibody of the invention or fragment thereof for the prevention,treatment or amelioration of one or more symptoms associated with a RSVinfection. Preferably, novel antibodies of the invention or fragmentsthereof are administered by pulmonary administration.

[0218] The invention also provides that an antibody or fragment thereofis packaged in a hermetically sealed container such as an ampoule orsachette indicating the quantity of antibody or antibody fragment. Inone embodiment, the antibody or antibody fragment is supplied as a drysterilized lyophilized powder or water free concentrate in ahermetically sealed container and can be reconstituted, e.g., with wateror saline to the appropriate concentration for administration to asubject. Preferably, the antibody or antibody fragment is supplied as adry sterile lyophilized powder in a hermetically sealed container at aunit dosage of at least 5 mg, more preferably at least 10 mg, at least15 mg, at least 25 mg, at least 35 mg, at least 45 mg, at least 50 mg,or at least 75 mg. The lyophilized antibody or antibody fragment shouldbe stored at between 2 and 8° C. in its original container and theantibody or antibody fragment should be administered within 12 hours,preferably within 6 hours, within 5 hours, within 3 hours, or within 1hour after being reconstituted. In an alternative embodiment, anantibody or fragment thereof is supplied in liquid form in ahermetically sealed container indicating the quantity and concentrationof the antibody or antibody fragment. Preferably, the liquid form of theantibody or fragment thereof is supplied in a hermetically sealedcontainer at least 1 mg/ml, more preferably at least 2.5 mg/ml, at least5 mg/ml, at least 8 mg/ml, at least 10 mg/ml, at least 15 mg/kg, or atleast 25 mg/ml.

[0219] In a specific embodiment, it may be desirable to administer thepharmaceutical compositions of the invention locally to the area in needof treatment; this may be achieved by, for example, and not by way oflimitation, local infusion, by injection, or by means of an implant,said implant being of a porous, non-porous, or gelatinous material,including membranes, such as sialastic membranes, or fibers. Preferably,when administering a an antibody of the invention or fragment thereof,care must be taken to use materials to which the antibody or antibodyfragment does not absorb.

[0220] In another embodiment, the composition can be delivered in avesicle, in particular a liposome (see Langer, Science 249:1527-1533(1990); Treat et al., in Liposomes in the Therapy of Infectious Diseaseand Cancer, Lopez-Berestein and Fidler (eds.), Liss, N.Y., pp. 353- 365(1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid.).

[0221] In yet another embodiment, the composition can be delivered in acontrolled release or sustained release system. In one embodiment, apump may be used to achieve controlled or sustained release (see Langer,supra; Sefton, 1987, CRC Crit. Ref. Biomed. Eng. 14:20; Buchwald et al.,1980, Surgery 88:507; Saudek et al., 1989, N. Engl. J. Med. 321:574). Inanother embodiment, polymeric materials can be used to achievecontrolled or sustained release of the antibodies of the invention orfragments thereof (see e.g., Medical Applications of Controlled Release,Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); ControlledDrug Bioavailability, Drug Product Design and Performance, Smolen andBall (eds.), Wiley, N.Y. (1984); Ranger and Peppas, 1983, J., Macromol.Sci. Rev. Macromol. Chem. 23:61; see also Levy et al., 1985, Science228:190; During et al., 1989, Ann. Neurol. 25:351; Howard et al., 1989,J. Neurosurg. 71:105); U.S. Pat. No. 5,679,377; U.S. Pat. No. 5,916,597;U.S. Pat. No. 5,912,015; U.S. Pat. No. 5,989,463; U.S. Pat. No.5,128,326; PCT Publication No. WO 99/15154; and PCT Publication No. WO99/20253. Examples of polymers used in sustained release formulationsinclude, but are not limited to, poly(2-hydroxy ethyl methacrylate),poly(methyl methacrylate), poly(acrylic acid), poly(ethylene-co-vinylacetate), poly(methacrylic acid), polyglycolides (PLG), polyanhydrides,poly(N-vinyl pyrrolidone), poly(vinyl alcohol), polyacrylamide,poly(ethylene glycol), polylactides (PLA), poly(lactide-co-glycolides)(PLGA), and polyorthoesters. In a preferred embodiment, the polymer usedin a sustained release formulation is inert, free of leachableimpurities, stable on storage, sterile, and biodegradable. In yetanother embodiment, a controlled or sustained release system can beplaced in proximity of the therapeutic target, i.e., the lungs, thusrequiring only a fraction of the systemic dose (see, e.g., Goodson, inMedical Applications of Controlled Release, supra, vol. 2, pp. 115-138(1984)).

[0222] Controlled release systems are discussed in the review by Langer(1990, Science 249:1527-1533). Any technique known to one of skill inthe art can be used to produce sustained release formulations comprisingone or more antibodies of the invention or fragments thereof. See, e.g.,U.S. Pat. No. 4,526,938, PCT publication WO 91/05548, PCT publication WO96/20698,. Ning et al., 1996, “Intratumoral Radioimmunotheraphy of aHuman Colon Cancer Xenograft Using a Sustained-Release Gel,”Radiotherapy & Oncology 39:179-189,. Song et al., 1995, “AntibodyMediated Lung Targeting of Long-Circulating Emulsions,” PDA Journal ofPharmaceutical Science & Technology 50:372-397, Cleek et al., 1997,“Biodegradable Polymeric Carriers for a bFGF Antibody for CardiovascularApplication,” Pro. Int'l. Symp. Control. Rel. Bioact. Mater. 24:853-854,and Lam et al., 1997, “Microencapsulation of Recombinant HumanizedMonoclonal Antibody for Local Delivery,” Proc. Int'l. Symp. Control Rel.Bioact. Mater. 24:759-760, each of which is incorporated herein byreference in their entirety.

[0223] In a specific embodiment where the composition of the inventionis a nucleic acid encoding an antibody or antibody fragment, the nucleicacid can be administered in vivo to promote expression of its encodedantibody or antibody fragment, by constructing it as part of anappropriate nucleic acid expression vector and administering it so thatit becomes intracellular, e.g., by use of a retroviral vector (see U.S.Pat. No. 4,980,286), or by direct injection, or by use of microparticlebombardment (e.g., a gene gun; Biolistic, Dupont), or coating withlipids or cell-surface receptors or transfecting agents, or byadministering it in linkage to a homeobox-like peptide which is known toenter the nucleus (see e.g., Joliot et al., 1991, Proc. Natl. Acad. Sci.USA 88:1864-1868), etc. Alternatively, a nucleic acid can be introducedintracellularly and incorporated within host cell DNA for expression byhomologous recombination.

[0224] The present invention also provides pharmaceutical compositions.Such compositions comprise a prophylactically or therapeuticallyeffective amount of an antibody or a fragment thereof, and apharmaceutically acceptable carrier. In a specific embodiment, the term“pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopeia orother generally recognized pharmacopeia for use in animals, and moreparticularly in humans. The term “carrier” refers to a diluent, adjuvant(e.g., Freund's adjuvant (complete and incomplete)), excipient, orvehicle with which the therapeutic is administered. Such pharmaceuticalcarriers can be sterile liquids, such as water and oils, including thoseof petroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. Water is a preferredcarrier when the pharmaceutical composition is administeredintravenously. Saline solutions and aqueous dextrose and glycerolsolutions can also be employed as liquid carriers, particularly forinjectable solutions. Suitable pharmaceutical excipients include starch,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, sodium stearate, glycerol monostearate, talc, sodium chloride,dried skim milk, glycerol, propylene, glycol, water, ethanol and thelike. The composition, if desired, can also contain minor amounts ofwetting or emulsifying agents, or pH buffering agents. Thesecompositions can take the form of solutions, suspensions, emulsion,tablets, pills, capsules, powders, sustained-release formulations andthe like. Oral formulation can include standard carriers such aspharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharine, cellulose, magnesium carbonate, etc. Examples ofsuitable pharmaceutical carriers are described in “Remington'sPharmaceutical Sciences” by E. W. Martin. Such compositions will containa prophylactically or therapeutically effective amount of the antibodyor fragment thereof, preferably in purified form, together with asuitable amount of carrier so as to provide the form for properadministration to the patient. The formulation should suit the mode ofadministration.

[0225] In a preferred embodiment, the composition is formulated inaccordance with routine procedures as a pharmaceutical compositionadapted for intravenous administration to human beings. Typically,compositions for intravenous administration are solutions in sterileisotonic aqueous buffer. Where necessary, the composition may alsoinclude a solubilizing agent and a local anesthetic such as lignocamneto ease pain at the site of the injection.

[0226] Generally, the ingredients of compositions of the invention aresupplied either separately or mixed together in unit dosage form, forexample, as a dry lyophilized powder or water free concentrate in ahermetically sealed container such as an ampoule or sachette indicatingthe quantity of active agent. Where the composition is to beadministered by infusion, it can be dispensed with an infusion bottlecontaining sterile pharmaceutical grade water or saline. Where thecomposition is administered by injection, an ampoule of sterile waterfor injection or saline can be provided so that the ingredients may bemixed prior to administration.

[0227] The compositions of the invention can be formulated as neutral orsalt forms. Pharmaceutically acceptable salts include those formed withanions such as those derived from hydrochloric, phosphoric, acetic,oxalic, tartaric acids, etc., and those formed with cations such asthose derived from sodium, potassium, ammonium, calcium, ferrichydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol,histidine, procaine, etc.

[0228] The amount of the composition of the invention which will beeffective in the treatment, prevention or amelioration of one or moresymptoms associated with a RSV infection can be determined by standardclinical techniques. For example, the dosage of the composition whichwill be effective in the treatment, prevention or amelioration of one ormore symptoms associated with a RSV infection can be determined byadministering the composition to a cotton rat, measuring the RSV titerafter challenging the cotton rat with 10⁵ pfu of RSV and comparing theRSV titer to that obtain for a cotton rat not administered thecomposition. Accordingly, a dosage that results in a 2 log decrease or a99% reduction in RSV titer in the cotton rat challenged with 10⁵ pfu ofRSV relative to the cotton rat challenged with 10⁵ pfu of RSV but notadministered the composition is the dosage of the composition that canbe administered to a human for the treatment, prevention or ameliorationof symptoms associated with RSV infection. The dosage of the compositionwhich will be effective in the treatment, prevention or amelioration ofone or more symptoms associated with a RSV infection can be determinedby administering the composition to an animal model (e.g. a cotton rator monkey) and measuring the serum titer of antibodies or fragmentsthereof that immunospecifically bind to a RSV antigen. Accordingly, adosage of the composition that results in a serum titer of at least 1μg/ml, preferably 2 μg/ml, 5 μg/ml, 10 μg/ml, 20 μg/ml, 25 μg/ml, atleast 35 μg/ml, at least 40 μg/ml, at least 50 μg/ml, at least 75 μg/ml,at least 100 ,g/ml, at least 125 μg/ml, at least 150 μg/ml, at least 200μg/ml, at least 250 μg/ml, at least 300 μg/ml, at least 350 μg/ml, atleast 400 μg/ml, or at least 450 μg/ml can be administered to a humanfor the treatment, prevention or amelioration of one or more symptomsassociated with RSV infection. In addition, in vitro assays mayoptionally be employed to help identify optimal dosage ranges.

[0229] The precise dose to be employed in the formulation will alsodepend on the route of administration, and the seriousness of the RSVinfection, and should be decided according to the judgment of thepractitioner and each patient's circumstances. Effective doses may beextrapolated from dose-response curves derived from in vitro or animalmodel (e.g., the cotton rat or Cynomolgous monkey) test systems.

[0230] For antibodies, the dosage administered to a patient is typically0.1 mg/kg to 100 mg/kg of the patient's body weight. Preferably, thedosage administered to a patient is between 0.1 mg/kg and 20 mg/kg ofthe patient's body weight, more preferably 1 mg/kg to 10 mg/kg of thepatient's body weight. Generally, human antibodies have a longerhalf-life within the human body than antibodies from other species dueto the immune response to the foreign polypeptides. Thus, lower dosagesof human antibodies and less frequent administration is often possible.Further, the dosage and frequency of administration of antibodies of theinvention or fragments thereof may be reduced by enhancing uptake andtissue penetration (e.g., into the lung) of the antibodies bymodifications such as, for example, lipidation.

[0231] In a specific embodiment, antibodies of the invention orfragments thereof, or compositions comprising antibodies of theinvention or fragments thereof are administered once a month just priorto or during the RSV season. In another embodiment, antibodies of theinvention or fragments thereof, or compositions comprising antibodies ofthe invention or fragments thereof are administered every two monthsjust prior to or during the RSV season. In yet another embodiment,antibodies of the invention or fragments thereof, or compositionscomprising antibodies of the invention or fragments thereof areadministered once just prior to or during the RSV season. The term “RSVseason” refers to the season when RSV infection is most likely to occur.Typically, the RSV season in the northern hemisphere commences inNovember and lasts through April.

[0232] In one embodiment, approximately 5 mg/kg or less (preferably 1.5mg/kg or less) of an antibody or fragment thereof whichimmunospecifically binds to a RSV antigen with a higher avidity and/orhigher affinity than previously known antibodies such as, e.g.,SYNAGIS®, is administered five times, 3 times, or 1 to 2 times during aRSV season to a mammal, preferably a human. In another embodiment,approximately 1.5 mg/kg of an antibody or a fragment thereof whichimmunospecifically binds to a RSV antigen with a higher avidity and/or ahigher affinity than known antibodies such as, e.g., SYNAGIS®, isadministered monthly five times during an RSV season to a mammal,preferably a human, intramuscularly. In another embodiment, 3 mg/kg ofan antibody or a fragment thereof which immunospecifically binds to aRSV antigen with a higher avidity and/or a higher affinity than knownantibodies such as, e.g., SYNAGIS® is administered monthly three timesduring an RSV season to a mammal, preferably a human, intramuscularly.In yet another embodiment, 5 mg/kg of an antibody or a fragment thereofwhich immunospecifically binds to a RSV antigen with a higher avidityand/or a higher affinity than known antibodies such as, e.g., SYNAGIS®is administered monthly one to two times during an RSV season to amammal, preferably a human, intramuscularly.

[0233] In a specific embodiment, 15 mg/kg of HL-SYNAGIS or anantigen-binding fragment thereof is administered to a mammal, preferablya human, intramuscularly five times during an RSV season. In anotherembodiment, approximately 5 mg/kg or less (preferably 1.5 mg/kg or less)of an antibody or fragment thereof which immunospecifically binds to aRSV antigen with a higher avidity and/or higher affinity than previouslyknown antibodies such as, e.g., SYNAGIS®, is administered five times, 3times, or 1 to 2 times during a RSV season to a mammal, preferably ahuman. In another embodiment, 3 mg/kg of antibody or a fragment thereofwhich immunospecifically binds to a RSV antigen with a higher avidityand/or a higher affinity known antibodies such as, e.g., SYNAGIS® andwhich has an increased in vivo half-life is administered monthly threetimes during an RSV season to a mammal, preferably a human,intramuscularly. In another embodiment, 5 mg/kg of antibody or afragment thereof which immunospecifically binds to a RSV antigen with ahigher avidity and/or a higher affinity than known antibodies such as,e.g., SYNAGIS® and which has an increased in vivo half-life isadministered to a mammal, preferably a human, intramuscularly twicetimes during an RSV season.

[0234] In a specific embodiment, an approximately 15 mg/kg bolus ofSYNAGIS® or an antigen-binding fragment thereof not in a sustainedrelease formulation is administered to a mammal, preferably a human, andafter a certain period of time less than 15 mg/kg (preferably 5 mg/kg orless, more preferably 3 mg/kg or less, and most preferably 1.5 mg/kg orless) of SYNAGIS® or an antibody fragment in a sustained release isadministered to said mammal intramuscularly two, three or four timesduring an RSV season. In accordance with this embodiment, a certainperiod of time can be 1 to 5 days, a week, two weeks, or a month. Inanother embodiment, approximately 15 mg/kg or less (preferably at least2 mg/kg, at least 5 mg/kg, or at least 10 mg/kg) of SYNAGIS® or anantigen-binding fragment thereof in a sustained release formulation isadministered to a mammal, preferably a human, intramuscularly two, threeor four times during an RSV season.

[0235] In another embodiment, approximately 15 mg/kg or less (preferablyat least 2 mg/kg, at least 5 mg/kg, or at least 10 mg/kg) of one or moreantibodies or fragments thereof which immunospecifically bind to one ormore RSV antigens is administered to the lungs of a mammal by pulmonarydelivery and then after a certain period of time (e.g., 15 minutes, 30minutes, 45 minutes, 1 hour, 6 hours, 12 hours, 1 day, 5 days, 10 days,20 days, 25 days, 30 days, or 40 days) approximately 15 mg/kg or less ofone or more said antibodies or antibody fragments is administeredintramusclarly said mammal. In another embodiment, approximately 15mg/kg or less (preferably at least 2 mg/kg, at least 5 mg/kg, or atleast 10 mg/kg) of one or more antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens is administered to amammal intramusclarly and then after a certain period of time (e.g., 15minutes, 30 minutes, 45 minutes, 1 hour, 6 hours, 12 hours, 1 day, 5days, 10 days, 20 days, 25 days, 30 days, or 40 days) approximately 15mg/kg or less of one or more said antibodies or antibody fragments isadministered to the lungs of said mammal.

[0236] 5.3.1. Gene Therapy

[0237] In a specific embodiment, nucleic acids comprising sequencesencoding antibodies or functional derivatives thereof, are administeredto treat, prevent or ameliorate one or more symptoms associated with RSVinfection, by way of gene therapy. Gene therapy refers to therapyperformed by the administration to a subject of an expressed orexpressible nucleic acid. In this embodiment of the invention, thenucleic acids produce their encoded antibody or antibody fragment thatmediates a prophylactic or therapeutic effect.

[0238] Any of the methods for gene therapy available in the art can beused according to the present invention. Exemplary methods are describedbelow.

[0239] For general reviews of the methods of gene therapy, see Goldspielet al., 1993, Clinical Pharmacy 12:488-505; Wu and Wu, 1991, Biotherapy3:87-95; Tolstoshev, 1993, Ann. Rev. Pharmacol. Toxicol. 32:573-596;Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, 1993,Ann. Rev. Biochem. 62:191-217; May, 1993, TIBTECH 11(5):155-215. Methodscommonly known in the art of recombinant DNA technology which can beused are described in Ausubel et al. (eds.), Current Protocols inMolecular Biology, John Wiley & Sons, NY (1993); and Kriegler, GeneTransfer and Expression, A Laboratory Manual, Stockton Press, NY (1990).

[0240] In a preferred aspect, a composition of the invention comprisesnucleic acids encoding an antibody, said nucleic acids being part of anexpression vector that expresses the antibody or fragments or chimericproteins or heavy or light chains thereof in a suitable host. Inparticular, such nucleic acids have promoters, preferably heterologouspromoters, operably linked to the antibody coding region, said promoterbeing inducible or constitutive, and, optionally, tissue-specific. Inanother particular embodiment, nucleic acid molecules are used in whichthe antibody coding sequences and any other desired sequences areflanked by regions that promote homologous recombination at a desiredsite in the genome, thus providing for intrachromosomal expression ofthe antibody encoding nucleic acids (Koller and Smithies, 1989, Proc.Natl. Acad. Sci. USA 86:8932-8935; Zijlstra et al., 1989, Nature342:435-438). In specific embodiments, the expressed antibody moleculeis a single chain antibody; alternatively, the nucleic acid sequencesinclude sequences encoding both the heavy and light chains, or fragmentsthereof, of the antibody.

[0241] Delivery of the nucleic acids into a subject may be eitherdirect, in which case the subject is directly exposed to the nucleicacid or nucleic acid-carrying vectors, or indirect, in which case, cellsare first transformed with the nucleic acids in vitro, then transplantedinto the subject. These two approaches are known, respectively, as invivo or ex vivo gene therapy.

[0242] In a specific embodiment, the nucleic acid sequences are directlyadministered in vivo, where it is expressed to produce the encodedproduct. This can be accomplished by any of numerous methods known inthe art, e.g., by constructing them as part of an appropriate nucleicacid expression vector and administering it so that they becomeintracellular, e.g., by infection using defective or attenuatedretrovirals or other viral vectors (see U.S. Pat. No. 4,980,286), or bydirect injection of naked DNA, or by use of microparticle bombardment(e.g., a gene gun; Biolistic, Dupont), or coating with lipids orcell-surface receptors or transfecting agents, encapsulation inliposomes, microparticles, or microcapsules, or by administering them inlinkage to a peptide which is known to enter the nucleus, byadministering it in linkage to a ligand subject to receptor-mediatedendocytosis (see, e.g., Wu and Wu, 1987, J. Biol. Chem. 262:4429-4432)(which can be used to target cell types specifically expressing thereceptors), etc. In another embodiment, nucleic acid-ligand complexescan be formed in which the ligand comprises a fusogenic viral peptide todisrupt endosomes, allowing the nucleic acid to avoid lysosomaldegradation. In yet another embodiment, the nucleic acid can be targetedin vivo for cell specific uptake and expression, by targeting a specificreceptor (see, e.g., PCT Publications WO 92/06180; WO 92/22635; WO92/20316; WO93/14188, WO 93/20221). Alternatively, the nucleic acid can beintroduced intracellularly and incorporated within host cell DNA forexpression, by homologous recombination (Koller and Smithies, 1989,Proc. Natl. Acad. Sci. USA 86:8932-8935; and Zijlstra et al., 1989,Nature 342:435-438).

[0243] In a specific embodiment, viral vectors that contains nucleicacid sequences encoding an antibody of the invention or fragmentsthereof are used. For example, a retroviral vector can be used (seeMiller et al., 1993, Meth. Enzymol. 217:581-599). These retroviralvectors contain the components necessary for the correct packaging ofthe viral genome and integration into the host cell DNA. The nucleicacid sequences encoding the antibody to be used in gene therapy arecloned into one or more vectors, which facilitates delivery of the geneinto a subject. More detail about retroviral vectors can be found inBoesen et al., 1994, Biotherapy 6:291-302, which describes the use of aretroviral vector to deliver the mdr 1 gene to hematopoietic stem cellsin order to make the stem cells more resistant to chemotherapy. Otherreferences illustrating the use of retroviral vectors in gene therapyare: Clowes et al., 1994, J. Clin. Invest. 93:644-651; Klein et al.,1994, Blood 83:1467-1473; Salmons and Gunzberg, 1993, Human Gene Therapy4:129-141; and Grossman and Wilson, 1993, Curr. Opin. in Genetics andDevel. 3:110-114.

[0244] Adenoviruses are other viral vectors that can be used in genetherapy. Adenoviruses are especially attractive vehicles for deliveringgenes to respiratory epithelia. Adenoviruses naturally infectrespiratory epithelia where they cause a mild disease. Other targets foradenovirus-based delivery systems are liver, the central nervous system,endothelial cells, and muscle. Adenoviruses have the advantage of beingcapable of infecting non-dividing cells. Kozarsky and Wilson, 1993,Current Opinion in Genetics and Development 3:499-503 present a reviewof adenovirus-based gene therapy. Bout et al., 1994, Human Gene Therapy5:3 -10 demonstrated the use of adenovirus vectors to transfer genes tothe respiratory epithelia of rhesus monkeys. Other instances of the useof adenoviruses in gene therapy can be found in Rosenfeld et al., 1991,Science 252:431-434; Rosenfeld et al., 1992, Cell 68:143-155;Mastrangeli et al., 1993, J. Clin. Invest. 91:225-234; PCT PublicationWO94/12649; and Wang et al., 1995, Gene Therapy 2:775-783. In apreferred embodiment, adenovirus vectors are used.

[0245] Adeno-associated virus (AAV) has also been proposed for use ingene therapy (Walsh et al., 1993, Proc. Soc. Exp. Biol. Med.204:289-300; and U.S. Pat. No. 5,436,146).

[0246] Another approach to gene therapy involves transferring a gene tocells in tissue culture by such methods as electroporation, lipofection,calcium phosphate mediated transfection, or viral infection. Usually,the method of transfer includes the transfer of a selectable marker tothe cells. The cells are then placed under selection to isolate thosecells that have taken up and are expressing the transferred gene. Thosecells are then delivered to a subject.

[0247] In this embodiment, the nucleic acid is introduced into a cellprior to administration in vivo of the resulting recombinant cell. Suchintroduction can be carried out by any method known in the art,including but not limited to transfection, electroporation,microinjection, infection with a viral or bacteriophage vectorcontaining the nucleic acid sequences, cell fusion, chromosome-mediatedgene transfer, microcellmediated gene transfer, spheroplast fusion, etc.Numerous techniques are known in the art for the introduction of foreigngenes into cells (see, e.g., Loeffler and Behr, 1993, Meth. Enzymol.217:599-618; Cohen et al., 1993, Meth. Enzymol. 217:618-644; Clin.Pharma. Ther. 29:69-92 (1985)) and may be used in accordance with thepresent invention, provided that the necessary developmental andphysiological functions of the recipient cells are not disrupted. Thetechnique should provide for the stable transfer of the nucleic acid tothe cell, so that the nucleic acid is expressible by the cell andpreferably heritable and expressible by its cell progeny.

[0248] The resulting recombinant cells can be delivered to a subject byvarious methods known in the art. Recombinant blood cells (e.g.,hematopoietic stem or progenitor cells) are preferably administeredintravenously. The amount of cells envisioned for use depends on thedesired effect, patient state, etc., and can be determined by oneskilled in the art.

[0249] Cells into which a nucleic acid can be introduced for purposes ofgene therapy encompass any desired, available cell type, and include butare not limited to epithelial cells, endothelial cells, keratinocytes,fibroblasts, muscle cells, hepatocytes; blood cells such as Tlymphocytes, B lymphocytes, monocytes, macrophages, neutrophils,eosinophils, megakaryocytes, granulocytes; various stem or progenitorcells, in particular hematopoietic stem or progenitor cells, e.g., asobtained from bone marrow, umbilical cord blood, peripheral blood, fetalliver, etc.

[0250] In a preferred embodiment, the cell used for gene therapy isautologous to the subject.

[0251] In an embodiment in which recombinant cells are used in genetherapy, nucleic acid sequences encoding an antibody or fragment thereofare introduced into the cells such that they are expressible by thecells or their progeny, and the recombinant cells are then administeredin vivo for therapeutic effect. In a specific embodiment, stem orprogenitor cells are used. Any stem and/or progenitor cells which can beisolated and maintained in vitro can potentially be used in accordancewith this embodiment of the present invention (see e.g., PCT PublicationWO 94/08598; Stemple and Anderson, 1992, Cell 7 1:973-985; Rheinwald,1980, Meth. Cell Bio. 21A:229; and Pittelkow and Scott, 1986, MayoClinic Proc. 61:771).

[0252] In a specific embodiment, the nucleic acid to be introduced forpurposes of gene therapy comprises an inducible promoter operably linkedto the coding region, such that expression of the nucleic acid iscontrollable by controlling the presence or absence of the appropriateinducer of transcription.

[0253] 5.4. Antibody Characterization and Demonstration of Therapeuticor Prophylactic Utility

[0254] Antibodies of the present invention or fragments thereof may becharacterized in a variety of ways. In particular, antibodies of theinvention or fragments thereof may be assayed for the ability toimmunospecifically bind to a RSV antigen. Such an assay may be performedin solution (e.g., Houghten, 1992, Bio/Techniques 13:412-421), on beads(Lam, 30 1991, Nature 354:82-84), on chips (Fodor, 1993, Nature364:555-556), on bacteria (U.S. Pat. No. 5,223,409), on spores (U.S.Pat. Nos. 5,571,698; 5,403,484; and 5,223,409), on plasmids (Cull etal., 1992, Proc. Natl. Acad. Sci. USA 89:1865-1869) or on phage (Scottand Smith, 1990, Science 249:386-390; Devlin, 1990, Science 249:404-406;Cwirla et al., 1990, Proc. Natl. Acad. Sci. USA 87:6378-6382; andFelici, 1991, J. Mol. Biol. 222:301-310) (each of these references isincorporated herein in its entirety by reference). Antibodies orfragments thereof that have been identified to immunospecifically bindto a RSV antigen or a fragment thereof can then be assayed for theirspecificity and affinity for a RSV antigen.

[0255] The antibodies of the invention or fragments thereof may beassayed for immunospecific binding to a RSV antigen and cross-reactivitywith other antigens by any method known in the art. Immunoassays whichcan be used to analyze immunospecific binding and cross-reactivityinclude, but are not limited to, competitive and non-competitive assaysystems using techniques such as western blots, radioimmunoassays, ELISA(enzyme linked immunosorbent assay), “sandwich” immunoassays,immunoprecipitation assays, precipitin reactions, gel diffusionprecipitin reactions, immunodiffusion assays, agglutination assays,complement-fixation assays, immunoradiometric assays, fluorescentimmunoassays, protein A immunoassays, to name but a few. Such assays areroutine and well known in the art (see, e.g., Ausubel et al, eds, 1994,Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc.,New York, which is incorporated by reference herein in its entirety).Exemplary immunoassays are described briefly below (but are not intendedby way of limitation).

[0256] Immunoprecipitation protocols generally comprise lysing apopulation of cells in a lysis buffer such as RIPA buffer (1% NP-40 orTriton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 0.01 Msodium phosphate at pH 7.2, 1% Trasylol) supplemented with proteinphosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin,sodium vanadate), adding the antibody of interest to the cell lysate,incubating for a period of time (e.g., 1 to 4 hours) at 40° C., addingprotein A and/or protein G sepharose beads to the cell lysate,incubating for about an hour or more at 40° C, washing the beads inlysis buffer and resuspending the beads in SDS/sample buffer. Theability of the antibody of interest to immunoprecipitate a particularantigen can be assessed by, e.g., western blot analysis. One of skill inthe art would be knowledgeable as to the parameters that can be modifiedto increase the binding of the antibody to an antigen and decrease thebackground (e.g., pre-clearing the cell lysate with sepharose beads).For further discussion regarding immunoprecipitation protocols see,e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology,Vol. 1, John Wiley & Sons, Inc., New York at 10.16.1.

[0257] Western blot analysis generally comprises preparing proteinsamples, electrophoresis of the protein samples in a polyacrylamide gel(e.g., 8%- 20% SDS-PAGE depending on the molecular weight of theantigen), transferring the protein sample from the polyacrylamide gel toa membrane such as nitrocellulose, PVDF or nylon, blocking the membranein blocking solution (e.g., PBS with 3% BSA or non-fat milk), washingthe membrane in washing buffer (e.g., PBS-Tween 20), blocking themembrane with primary antibody (the antibody of interest) diluted inblocking buffer, washing the membrane in washing buffer, blocking themembrane with a secondary antibody (which recognizes the primaryantibody, e.g., an anti-human antibody) conjugated to an enzymaticsubstrate (e.g., horseradish peroxidase or alkaline phosphatase) orradioactive molecule (e.g., ³²p or ¹²⁵I) diluted in blocking buffer,washing the membrane in wash buffer, and detecting the presence of theantigen. One of skill in the art would be knowledgeable as to theparameters that can be modified to increase the signal detected and toreduce the background noise. For further discussion regarding westernblot protocols see, e.g., Ausubel et al, eds, 1994, Current Protocols inMolecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 10.8.1.

[0258] ELISAs comprise preparing antigen, coating the well of a 96 wellmicrotiter plate with the antigen, adding the antibody of interestconjugated to a detectable compound such as an enzymatic substrate(e.g., horseradish peroxidase or alkaline phosphatase) to the well andincubating for a period of time, and detecting the presence of theantigen. In ELISAs the antibody of interest does not have to beconjugated to a detectable compound; instead, a second antibody (whichrecognizes the antibody of interest) conjugated to a detectable compoundmay be added to the well. Further, instead of coating the well with theantigen, the antibody may be coated to the well. In this case, a secondantibody conjugated to a detectable compound may be added following theaddition of the antigen of interest to the coated well. One of skill inthe art would be knowledgeable as to the parameters that can be modifiedto increase the signal detected as well as other variations of ELISAsknown in the art. For further discussion regarding ELISAs see, e.g.,Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol.1, John Wiley & Sons, Inc., New York at 11.2.1.

[0259] The binding affinity of an antibody to an antigen and theoff-rate of an antibody-antigen interaction can be determined bycompetitive binding assays. One example of a competitive binding assayis a radioimmunoassay comprising the incubation of labeled antigen(e.g., ³H or ¹²⁵I) with the antibody of interest in the presence ofincreasing amounts of unlabeled antigen, and the detection of theantibody bound to the labeled antigen. The affinity of the antibody ofthe present invention or a fragment thereof for a RSV antigen and thebinding off-rates can be determined from the data by scatchard plotanalysis. Competition with a second antibody can also be determinedusing radioimmunoassays. In this case, a RSV antigen is incubated withan antibody of the present invention or a fragment thereof conjugated toa labeled compound (e.g., ³H or ¹²⁵I) in the presence of increasingamounts of an unlabeled second antibody.

[0260] In a preferred embodiment, BlAcore kinetic analysis is used todetermine the binding on and off rates of antibodies or fragmentsthereof to a RSV antigen. BlAcore kinetic analysis comprises analyzingthe binding and dissociation of a RSV antigen from chips withimmobilized antibodies or fragments thereof on their surface (see theExample section infra).

[0261] The antibodies of the invention or fragments thereof can also beassayed for their ability to inhibit the binding of RSV to its host cellreceptor using techniques known to those of skill in the art. Forexample, cells expressing the receptor for RSV can be contacted with RSVin the presence or absence of an antibody or fragment thereof and theability of the antibody or fragment thereof to inhibit RSV's binding canmeasured by, for example, flow cytometry or a scintillation assay. RSV(e.g., a RSV antigen such as F glycoprotein or G glycoprotein) or theantibody or antibody fragment can be labeled with a detectable compoundsuch as a radioactive label (e.g., ³²P, ³⁵S, and ¹²⁵I) or a fluorescentlabel (e.g, fluorescein isothiocyanate, rhodamine, phycoerythrin,phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine) toenable detection of an interaction between RSV and its host cellreceptor. Alternatively, the ability of antibodies or fragments thereofto inhibit RSV from binding to its receptor can be determined incell-free assays. For example, RSV or a RSV antigen such as Gglycoprotein can be contacted with an antibody or fragment thereof andthe ability of the antibody or antibody fragment to inhibit RSV or theRSV antigen from binding to its host cell receptor can be determined.Preferably, the antibody or the antibody fragment is immobilized on asolid support and RSV or a RSV antigen is labeled with a detectablecompound. Alternatively, RSV or a RSV antigen is immobilized on a solidsupport and the antibody or fragment thereof is labeled with adetectable compound. RSV or a RSV antigen may be partially or completelypurified (e.g., partially or completely free of other polypeptides) orpart of a cell lysate. Further, an RSV antigen may be a fusion proteincomprising the RSV antigen and a domain such asglutathionine-S-transferase. Alternatively, an RSV antigen can bebiotinylated using techniques well known to those of skill in the art(e.g., biotinylation kit, Pierce Chemicals; Rockford, Ill.).

[0262] The antibodies of the invention or fragments thereof can also beassayed for their ability to inhibit or downregulate RSV replicationusing techniques known to those of skill in the art. For example, RSVreplication can be assayed by a plaque assay such as described, e.g., byJohnson et al., 1997, Journal of Infectious Diseases 176:1215-1224. Theantibodies of the invention or fragments thereof can also be assayed fortheir ability to inhibit or downregulate the expression of RSVpolypeptides. Techniques known to those of skill in the art, including,but not limited to, Western blot analysis, Northern blot analysis, andRT-PCR can be used to measure the expression of RSV polypeptides.Further, the antibodies of the invention or fragments thereof can beassayed for their ability to prevent the formation of syncytia.

[0263] The antibodies of the invention or fragments thereof arepreferably tested in vitro, and then in vivo for the desired therapeuticor prophylactic activity, prior to use in humans. For example, in vitroassays which can be used to determine whether administration of aspecific antibody or composition of the present invention is indicated,include in vitro cell culture assays in which a subject tissue sample isgrown in culture, and exposed to or otherwise administered an antibodyor composition of the present invention, and the effect of such anantibody or composition of the present invention upon the tissue sampleis observed. In various specific embodiments, in vitro assays can becarried out with representative cells of cell types involved in a RSVinfection (e.g., respiratory epithelial cells), to determine if anantibody or composition of the present invention has a desired effectupon such cell types. Preferably, the antibodies or compositions of theinvention are also tested in in vitro assays and animal model systemsprior to administration to humans. In a specific embodiment, cotton ratsare administered an antibody the invention or fragment thereof, or acomposition of the invention, challenged with 10⁵ pfu of RSV, and fouror more days later the rats are sacrificed and RSV titer and anti-RSVantibody serum titer is determined. Further, in accordance with thisembodiment, the tissues (e.g., the lung tissues) from the sacrificedrats can be examined for histological changes.

[0264] In accordance with the invention, clinical trials with humansubjects need not be performed in order to demonstrate the prophylacticand/or therapeutic efficacy of antibodies of the invention or fragmentsthereof. In vitro and animal model studies using the antibodies orfragments thereof can be extrapolated to humans and are sufficient fordemonstrating the prophylactic and/or therapeutic utility of saidantibodies or antibody fragments.

[0265] Antibodies or compositions of the present invention for use intherapy can be tested for their toxicity in suitable animal modelsystems, including but not limited to rats, mice, cows, monkeys, andrabbits. For in vivo testing of an antibody or composition's toxicityany animal model system known in the art may be used.

[0266] Efficacy in treating or preventing viral infection may bedemonstrated by detecting the ability of an antibody or composition ofthe invention to inhibit the replication of the virus, to inhibittransmission or prevent the virus from establishing itself in its host,to reduce the incidence of RSV infection, or to prevent, ameliorate oralleviate one or more symptoms associated with RSV infection. Thetreatment is considered therapeutic if there is, for example, areduction is viral load, amelioration of one or more symptoms, areduction in the duration of a RSV infection, or a decrease in mortalityand/or morbidity following administration of an antibody or compositionof the invention. Further, the treatment is considered therapeutic ifthere is an increase in the immune response following the administrationof one or more antibodies or fragments thereof which immunospecificallybind to one or more RSV antigens.

[0267] Antibodies or compositions of the invention can be tested invitro and in vivo for the ability to induce the expression of cytokinessuch as IFN-α, IFN-β, IFN-γ, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8,IL-9, IL-10, IL-12 and IL-15. Techniques known to those of skill in theart can be used to measure the level of expression of cytokines. Forexample, the level of expression of cytokines can be measured byanalyzing the level of RNA of cytokines by, for example, RT-PCR andNorthern blot analysis, and by analyzing the level of cytokines by, forexample, immunoprecipitation followed by western blot analysis andELISA. In a preferred embodiment, an antibody or composition of theinvention is tested for its ability to induce the expression of IFN-γ.

[0268] Antibodies or compositions of the invention can be tested invitro and in vivo for their ability to modulate the biological activityof immune cells, preferably human immune cells (e.g., T-cells, B-cells,and Natural Killer cells). The ability of an antibody or composition ofthe invention to modulate the biological activity of immune cells can beassessed by detecting the expression of antigens, detecting theproliferation of immune cells, detecting the activation of signalingmolecules, detecting the effector function of immune cells, or detectingthe differentiation of immune cells. Techniques known to those of skillin the art can be used for measuring these activities. For example,cellular proliferation can be assayed by ³H-thymidine incorporationassays and trypan blue cell counts. Antigen expression can be assayed,for example, by immunoassays including, but are not limited to,competitive and non-competitive assay systems using techniques such aswestern blots, immunohistochemistry radioimmunoassays, ELISA (enzymelinked immunosorbent assay), “sandwich” immunoassays,immunoprecipitation assays, precipitin reactions, gel diffusionprecipitin reactions, immunodiffusion assays, agglutination assays,complement-fixation assays, immunoradiometric assays, fluorescentimmunoassays, protein A immunoassays and FACS analysis. The activationof signaling molecules can be assayed, for example, by kinase assays andelectrophoretic shift assays (EMSAs).

[0269] Antibodies or compositions of the invention can also be testedfor their ability to inhibit viral replication or reduce viral load inin vitro, ex vivo and in vivo assays. Antibodies or compositions of theinvention can also be tested for their ability to decrease the timecourse of RSV infection. Antibodies or compositions of the invention canalso be tested for their ability to increase the survival period ofhumans suffering from RSV infection by at least 25%, preferably at least50%, at least 60%, at least 75%, at least 85%, at least 95%, or at least99%. Further, antibodies or compositions of the invention can be testedfor their ability reduce the hospitalization period of humans sufferingfrom RSV infection by at least 60%, preferably at least 75%, at least85%, at least 95%, or at least 99%. Techniques known to those of skillin the art can be used to analyze the function of the antibodies orcompositions of the invention in vivo.

[0270] 5.5. Diagnostic Uses of Antibodies

[0271] Labeled antibodies, fragments and derivatives and analogsthereof, which immunospecifically bind to a RSV antigen can be used fordiagnostic purposes to detect, diagnose, or monitor a RSV infection. Theinvention provides for the detection of a RSV infection, comprising: (a)assaying the expression of a RSV antigen in cells or a tissue sample ofa subject using one or more antibodies or fragments thereof thatimmunospecifically bind to the RSV antigen; and (b) comparing the levelof the RSV antigen with a control level, e.g., levels in normal tissuesamples not infected with RSV, whereby an increase in the assayed levelof RSV antigen compared to the control level of the RSV antigen isindicative of a RSV infection.

[0272] The invention provides a diagnostic assay for diagnosing a RSVinfection, comprising: (a) assaying for the level of a RSV antigen incells or a tissue sample of an individual using one or more antibodiesor fragments thereof that immunospecifically bind to a RSV antigen; and(b) comparing the level of the RSV antigen with a control level, e.g.,levels in normal tissue samples not infected with RSV, whereby anincrease in the assayed RSV antigen level compared to the control levelof the RSV antigen is indicative of a RSV infection. A more definitivediagnosis of RSV infection may allow health professionals to employpreventative measures or aggressive treatment earlier thereby preventingthe development or further progression of RSV infection.

[0273] Antibodies of the invention or fragments thereof can be used toassay RSV antigen levels in a biological sample using classicalimmunohistological methods as described herein or as known to those ofskill in the art (e.g., see Jalkanen et al., 1985, J. Cell. Biol.101:976-985; and Jalkanen et al., 1987, J. Cell . Biol. 105:3087-3096).Other antibody-based methods useful for detecting protein geneexpression include immunoassays, such as the enzyme linked immunosorbentassay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assaylabels are known in the art and include enzyme labels, such as, glucoseoxidase; radioisotopes, such as iodine (¹²⁵I, ¹²¹I), carbon (¹⁴C),sulfur (35S), tritium (³H), indium (¹²¹In), and technetium (⁹⁹Tc);luminescent labels, such as luminol; and fluorescent labels, such asfluorescein and rhodamine, and biotin.

[0274] One aspect of the invention is the detection and diagnosis of aRSV infection in a human. In one embodiment, diagnosis comprises: a)administering (for example, parenterally, subcutaneously, orintraperitoneally) to a subject an effective amount of a labeledantibody or fragment thereof that immunospecifically binds to a RSVantigen; b) waiting for a time interval following the administering forpermitting the labeled antibody or fragment thereof to preferentiallyconcentrate at sites in the subject (e.g., the lungs) where the RSVantigen is expressed (and for unbound labeled molecule to be cleared tobackground level); c) determining background level; and d) detecting thelabeled antibody or fragment thereof in the subject, such that detectionof labeled antibody or fragment thereof above the background levelindicates that the subject has a RSV infection. Background level can bedetermined by various methods including, comparing the amount of labeledmolecule detected to a standard value previously determined for aparticular system.

[0275] It will be understood in the art that the size of the subject andthe imaging system used will determine the quantity of imaging moietyneeded to produce diagnostic images. In the case of a radioisotopemoiety, for a human subject, the quantity of radioactivity injected willnormally range from about 5 to 20 millicuries of ⁹⁹Tc. The labeledantibody or antibody fragment will then preferentially accumulate at thelocation of cells which contain the specific protein. In vivo tumorimaging is described in S. W. Burchiel et al., “Immunopharmacokineticsof Radiolabeled Antibodies and Their Fragments.” (Chapter 13 in TumorImaging: The Radiochemical Detection of Cancer, S. W. Burchiel and B. A.Rhodes, eds., Masson Publishing Inc. (1982).

[0276] Depending on several variables, including the type of label usedand the mode of administration, the time interval following theadministration for permitting the labeled molecule to preferentiallyconcentrate at sites in the subject and for unbound labeled molecule tobe cleared to background level is 6 to 48 hours or 6 to 24 hours or 6 to12 hours. In another embodiment the time interval followingadministration is 5 to 20 days or 5 to 10 days.

[0277] In one embodiment, monitoring of a RSV infection is carried outby repeating the method for diagnosing the RSV infection, for example,one month after initial diagnosis, six months after initial diagnosis,one year after initial diagnosis, etc.

[0278] Presence of the labeled molecule can be detected in the subjectusing methods known in the art for in vivo scanning. These methodsdepend upon the type of label used. Skilled artisans will be able todetermine the appropriate method for detecting a particular label.Methods and devices that may be used in the diagnostic methods of theinvention include, but are not limited to, computed tomography (CT),whole body scan such as position emission tomography (PET), magneticresonance imaging (MRI), and sonography.

[0279] In a specific embodiment, the molecule is labeled with aradioisotope and is detected in the patient using a radiation responsivesurgical instrument (Thurston et al., U.S. Pat. No. 5,441,050). Inanother embodiment, the molecule is labeled with a fluorescent compoundand is detected in the patient using a fluorescence responsive scanninginstrument. In another embodiment, the molecule is labeled with apositron emitting metal and is detected in the patient using positronemission-tomography. In yet another embodiment, the molecule is labeledwith a paramagnetic label and is detected in a patient using magneticresonance imaging (MRI).

[0280] 5.6. Methods Producing Antibodies

[0281] The antibodies of the invention or fragments thereof can beproduced by any method known in the art for the synthesis of antibodies,in particular, by chemical synthesis or preferably, by recombinantexpression techniques.

[0282] Polyclonal antibodies to a RSV antigen can be produced by variousprocedures well known in the art. For example, a RSV antigen can beadministered to various host animals including, but not limited to,rabbits, mice, rats, etc. to induce the production of sera containingpolyclonal antibodies specific for the RSV antigen. Various adjuvantsmay be used to increase the immunological response, depending on thehost species, and include but are not limited to, Freund's (complete andincomplete), mineral gels such as aluminum hydroxide, surface activesubstances such as lysolecithin, pluronic polyols, polyanions, peptides,oil emulsions, keyhole limpet hemocyanins, dinitrophenol, andpotentially useful human adjuvants such as BCG (bacille Calmette-Guerin)and corynebacterium parvum. Such adjuvants are also well known in theart.

[0283] Monoclonal antibodies can be prepared using a wide variety oftechniques known in the art including the use of hybridoma, recombinant,and phage display technologies, or a combination thereof. For example,monoclonal antibodies can be produced using hybridoma techniquesincluding those known in the art and taught, for example, in Harlow etal., Antibodies: A Laboratory Manual, (Cold Spring Harbor LaboratoryPress, 2nd ed. 1988); Hammerling, et al., in: Monoclonal Antibodies andT-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981) (said referencesincorporated by reference in their entireties). The term “monoclonalantibody” as used herein is not limited to antibodies produced throughhybridoma technology. The term “monoclonal antibody” refers to anantibody that is derived from a single clone, including any eukaryotic,prokaryotic, or phage clone, and not the method by which it is produced.

[0284] Methods for producing and screening for specific antibodies usinghybridoma technology are routine and well known in the art. Briefly,mice can be immunized with a RSV antigen and once an immune response isdetected, e.g., antibodies specific for the RSV antigen are detected inthe mouse serum, the mouse spleen is harvested and splenocytes isolated.The splenocytes are then fused by well known techniques to any suitablemyeloma cells, for example cells from cell line SP20 available from theATCC. Hybridomas are selected and cloned by limited dilution. Thehybridoma clones are then assayed by methods known in the art for cellsthat secrete antibodies capable of binding a polypeptide of theinvention. Ascites fluid, which generally contains high levels ofantibodies, can be generated by immunizing mice with positive hybridomaclones.

[0285] Accordingly, the present invention provides methods of generatingmonoclonal antibodies as well as antibodies produced by the methodcomprising culturing a hybridoma cell secreting an antibody of theinvention wherein, preferably, the hybridoma is generated by fusingsplenocytes isolated from a mouse immunized with a RSV antigen withmyeloma cells and then screening the hybridomas resulting from thefusion for hybridoma clones that secrete an antibody able to bind a RSVantigen.

[0286] Antibody fragments which recognize specific RSV epitopes may begenerated by any technique known to those of skill in the art. Forexample, Fab and F(ab′)2 fragments of the invention may be produced byproteolytic cleavage of immunoglobulin molecules, using enzymes such aspapain (to produce Fab fragments) or pepsin (to produce F(ab′)2fragments). F(ab′)2 fragments contain the variable region, the lightchain constant region and the CH1 domain of the heavy chain. Further,the antibodies of the present invention can also be generated usingvarious phage display methods known in the art.

[0287] In phage display methods, functional antibody domains aredisplayed on the surface of phage particles which carry thepolynucleotide sequences encoding them. In particular, DNA sequencesencoding VH and VL domains are amplified from animal cDNA libraries(e.g., human or murine cDNA libraries of lymphoid tissues). The DNAencoding the VH and VL domains are recombined together with an scFvlinker by PCR and cloned into a phagemid vector (e.g., p CANTAB 6 orpComb 3 HSS). The vector is electroporated in E. coli and the E. coli isinfected with helper phage. Phage used in these methods are typicallyfilamentous phage including fd and M13 and the VH and VL domains areusually recombinantly fused to either the phage gene III or gene VIII.Phage expressing an antigen binding domain that binds to a RSV antigenof interest can be selected or identified with antigen, e.g., usinglabeled antigen or antigen bound or captured to a solid surface or bead.Examples of phage display methods that can be used to make theantibodies of the present invention include those disclosed in Brinkmanet al., 1995, J. Immunol. Methods 182:41-50; Ames et al., 1995, J.Immunol. Methods 184:177-186; Kettleborough et al., 1994, Eur. J.Immunol. 24:952-958; Persic et al., 1997, Gene 187:9-18; Burton et al.,1994, Advances in Immunology 57:191-280; PCT application No. PCT/GB91/O1134; PCT publication Nos. WO 90/02809, WO 91/10737, WO 92/01047, WO92/18619, WO 93/1 1236, WO 95/15982, WO 95/20401, and W097/13844; andU.S. Pat. Nos. 5,698,426, 5,223,409, 5,403,484, 5,580,717, 5,427,908,5,750,753, 5,821,047, 5,571,698, 5,427,908, 5,516,637, 5,780,225,5,658,727, 5,733,743 and 5,969,108; each of which is incorporated hereinby reference in its entirety.

[0288] As described in the above references, after phage selection, theantibody coding regions from the phage can be isolated and used togenerate whole antibodies, including human antibodies, or any otherdesired antigen binding fragment, and expressed in any desired host,including mammalian cells, insect cells, plant cells, yeast, andbacteria, e.g., as described below. Techniques to recombinantly produceFab, Fab′ and F(ab′)2 fragments can also be employed using methods knownin the art such as those disclosed in PCT publication No. WO 92/22324;Mullinax et al., 1992, BioTechniques 12(6):864-869; Sawai et al., 1995,AJRI 34:26-34; and Better et al., 1988, Science 240:1041-1043 (saidreferences incorporated by reference in their entireties).

[0289] To generate whole antibodies, PCR primers including VH or VLnucleotide sequences, a restriction site, and a flanking sequence toprotect the restriction site can be used to amplify the VH or VLsequences in scFv clones. Utilizing cloning techniques known to those ofskill in the art, the PCR amplified VH domains can be cloned intovectors expressing a VH constant region, e.g., the human gamma 4constant region, and the PCR amplified VL domains can be cloned intovectors expressing a VL constant region, e.g., human kappa or lambaconstant regions. Preferably, the vectors for expressing the VH or VLdomains comprise an EF-1α promoter, a secretion signal, a cloning sitefor the variable domain, constant domains, and a selection marker suchas neomycin. The VH and VL domains may also cloned into one vectorexpressing the necessary constant regions. The heavy chain conversionvectors and light chain conversion vectors are then co-transfected intocell lines to generate stable or transient cell lines that expressfull-length antibodies, e.g., IgG, using techniques known to those ofskill in the art.

[0290] For some uses, including in vivo use of antibodies in humans andin vitro detection assays, it may be preferable to use human or chimericantibodies. Completely human antibodies are particularly desirable fortherapeutic treatment of human subjects. Human antibodies can be made bya variety of methods known in the art including phage display methodsdescribed above using antibody libraries derived from humanimmunoglobulin sequences. See also U.S. Pat. Nos. 4,444,887 and4,716,111; and PCT publications WO 98/46645, WO 98/50433, WO 98/24893,W098/16654, WO 96/34096, WO 96/33735, and WO 91/10741; each of which isincorporated herein by reference in its entirety.

[0291] Human antibodies can also be produced using transgenic mice whichare incapable of expressing functional endogenous immunoglobulins, butwhich can express human immunoglobulin genes. For example, the humanheavy and light chain immunoglobulin gene complexes may be introducedrandomly or by homologous recombination into mouse embryonic stem cells.Alternatively, the human variable region, constant region, and diversityregion may be introduced into mouse embryonic stem cells in addition tothe human heavy and light chain genes. The mouse heavy and light chainimmunoglobulin genes may be rendered non-functional separately orsimultaneously with the introduction of human immunoglobulin loci byhomologous recombination. In particular, homozygous deletion of theJ_(H) region prevents endogenous antibody production. The modifiedembryonic stem cells are expanded and microinjected into blastocysts toproduce chimeric mice. The chimeric mice are then be bred to producehomozygous offspring which express human antibodies. The transgenic miceare immunized in the normal fashion with a selected antigen, e.g., allor a portion of a polypeptide of the invention. Monoclonal antibodiesdirected against the antigen can be obtained from the immunized,transgenic mice using conventional hybridoma technology. The humanimmunoglobulin transgenes harbored by the transgenic mice rearrangeduring B cell differentiation, and subsequently undergo class switchingand somatic mutation. Thus, using such a technique, it is possible toproduce therapeutically useful IgG, IgA, IgM and IgE antibodies. For anoverview of this technology for producing human antibodies, see Lonbergand Huszar (1995, Int. Rev. Immunol. 13:65-93). For a detaileddiscussion of this technology for producing human antibodies and humanmonoclonal antibodies and protocols for producing such antibodies, see,e.g., PCT publication Nos. WO 98/24893, WO 96/34096, and WO 96/33735;and U.S. Pat. Nos. 5,413,923, 5,625,126, 5,633,425, 5,569,825,5,661,016, 5,545,806, 5,814,318, and 5,939,598, which are incorporatedby reference herein in their entirety. In addition, companies such asAbgenix, Inc. (Freemont, Calif.) and Genpharm (San Jose, Caif.) can beengaged to provide human antibodies directed against a selected antigenusing technology similar to that described above.

[0292] A chimeric antibody is a molecule in which different portions ofthe antibody are derived from different immunoglobulin molecules such asantibodies having a variable region derived from a human antibody and anon-human immunoglobulin constant region. Methods for producing chimericantibodies are known in the art. See e.g., Morrison, 1985, Science229:1202; Oi et al., 1986, BioTechniques 4:214; Gillies et al., 1989, J.Immunol. Methods 125:191-202; and U.S. Patent Nos. 5,807,715, 4,816,567,and 4,816,397, which are incorporated herein by reference in theirentirety. Chimeric antibodies comprising one or more CDRs from humanspecies and framework regions from a non-human immunoglobulin moleculecan be produced using a variety of techniques known in the artincluding, for example, CDR-grafting (EP 239,400; PCT publication No. WO91/09967; and U.S. Pat. Nos. 5,225,539, 5,530,101, and 5,585,089),veneering or resurfacing (EP 592,106; EP 519,596; Padlan, 1991,Molecular Immunology 28(4/5):489-498; Studnicka et al., 1994, ProteinEngineering 7(6):805-814; and Roguska et al., 1994, PNAS 91:969-973),and chain shuffling (U.S. Pat. No. 5,565,332). In a preferredembodiment, chimeric antibodies comprise a human CDR3 having an aminoacid sequence of any one of the CDR3 listed in Table 2 and non-humanframework regions. Often, framework residues in the framework regionswill be substituted with the corresponding residue from the CDR donorantibody to alter, preferably improve, antigen binding. These frameworksubstitutions are identified by methods well known in the art, e.g., bymodeling of the interactions of the CDR and framework residues toidentify framework residues important for antigen binding and sequencecomparison to identify unusual framework residues at particularpositions. (See, e.g., Queen et al., U.S. Pat. No. 5,585,089; andRiechmann et al., 1988, Nature 332:323, which are incorporated herein byreference in their entireties.)

[0293] Further, the antibodies of the invention can, in turn, beutilized to generate anti-idiotype antibodies that “mimic” RSV antigensusing techniques well known to those skilled in the art. (See, e.g.,Greenspan & Bona, 1989, FASEB J. 7(5):437-444; and Nissinoff, 1991, J.Immunol. 147(8):2429-2438). For example, antibodies of the inventionwhich bind to and competitively inhibit the binding of RSV (asdetermined by assays well known in the art and disclosed in supra) toits host cell receptor can be used to generate anti-idiotypes that“mimic” a RSV antigen binding domain and, as a consequence, bind to andneutralize RSV and/or its host cell receptor. Such neutralizinganti-idiotypes or Fab fragments of such anti-idiotypes can be used intherapeutic regimens to neutralize RSV. For example, such anti-idiotypicantibodies can be used to bind RSV and/or to bind its host cellreceptors, and thereby block infection.

[0294] 5.6.1. Polynucleotides Encoding an Antibody

[0295] The invention provides polynucleotides comprising a nucleotidesequence encoding an antibody of the invention or a fragment thereof.The invention also encompasses polynucleotides that hybridize under highstringency, intermediate or lower stringency hybridization conditions,e.g., as defined supra, to polynucleotides that encode an antibody ofthe invention.

[0296] The polynucleotides may be obtained, and the nucleotide sequenceof the polynucleotides determined, by any method known in the art. Sincethe amino acid sequences of the antibodies are known (as described inTable 2), nucleotide sequences encoding these antibodies can bedetermined using methods well known in the art, i.e., nucleotide codonsknown to encode particular amino acids are assembled in such a way togenerate a nucleic acid that encodes the antibody or fragment thereof ofthe invention. Such a polynucleotide encoding the antibody may beassembled from chemically synthesized oligonucleotides (e.g., asdescribed in Kutmeier et al., 1994, BioTechniques 17:242), which,briefly, involves the synthesis of overlapping oligonucleotidescontaining portions of the sequence encoding the antibody, annealing andligating of those oligonucleotides, and then amplification of theligated oligonucleotides by PCR.

[0297] Alternatively, a polynucleotide encoding an antibody may begenerated from nucleic acid from a suitable source. If a clonecontaining a nucleic acid encoding a particular antibody is notavailable, but the sequence of the antibody molecule is known, a nucleicacid encoding the immunoglobulin may be chemically synthesized orobtained from a suitable source (e.g., an antibody cDNA library, or acDNA library generated from, or nucleic acid, preferably poly A+RNA,isolated from, any tissue or cells expressing the antibody, such ashybridoma cells selected to express an antibody of the invention) by PCRamplification using synthetic primers hybridizable to the 3′ and 5 ′ends of the sequence or by cloning using an oligonucleotide probespecific for the particular gene sequence to identify, e.g., a cDNAclone from a cDNA library that encodes the antibody. Amplified nucleicacids generated by PCR may then be cloned into replicable cloningvectors using any method well known in the art.

[0298] Once the nucleotide sequence of the antibody is determined, thenucleotide sequence of the antibody may be manipulated using methodswell known in the art for the manipulation of nucleotide sequences,e.g., recombinant DNA techniques, site directed mutagenesis, PCR, etc.(see, for example, the techniques described in Sambrook et al., 1990,Molecular Cloning, A Laboratory Manual, 2d Ed., Cold Spring HarborLaboratory, Cold Spring Harbor, N.Y. and Ausubel et al., eds., 1998,Current Protocols in Molecular Biology, John Wiley & Sons, NY, which areboth incorporated by reference herein in their entireties), to generateantibodies having a different amino acid sequence, for example to createamino acid substitutions, deletions, and/or insertions.

[0299] In a specific embodiment, one or more of the CDRs is insertedwithin framework regions using routine recombinant DNA techniques. Theframework regions may be naturally occurring or consensus frameworkregions, and preferably human framework regions (see, e.g., Chothia etal., 1998, J. Mol. Biol. 278: 457-479 for a listing of human frameworkregions). Preferably, the polynucleotide generated by the combination ofthe framework regions and CDRs encodes an antibody that specificallybinds to a RSV antigen. Preferably, as discussed supra, one or moreamino acid substitutions may be made within the framework regions, and,preferably, the amino acid substitutions improve binding of the antibodyto its antigen. Additionally, such methods may be used to make aminoacid substitutions or deletions of one or more variable region cysteineresidues participating in an intrachain disulfide bond to generateantibody molecules lacking one or more intrachain disulfide bonds. Otheralterations to the polynucleotide are encompassed by the presentinvention and within the skill of the art.

[0300] 5.6.2. Recombinant Expression of an Antibody

[0301] Recombinant expression of an antibody of the invention,derivative or analog thereof, (e.g., a heavy or light chain of anantibody of the invention or a portion thereof or a single chainantibody of the invention), requires construction of an expressionvector containing a polynucleotide that encodes the antibody. Once apolynucleotide encoding an antibody molecule or a heavy or light chainof an antibody, or portion thereof (preferably, but not necessarily,containing the heavy or light chain variable domain), of the inventionhas been obtained, the vector for the production of the antibodymolecule may be produced by recombinant DNA technology using techniqueswell known in the art. Thus, methods for preparing a protein byexpressing a polynucleotide containing an antibody encoding nucleotidesequence are described herein. Methods which are well known to thoseskilled in the art can be used to construct expression vectorscontaining antibody coding sequences and appropriate transcriptional andtranslational control signals. These methods include, for example, invitro recombinant DNA techniques, synthetic techniques, and in vivogenetic recombination. The invention, thus, provides replicable vectorscomprising a nucleotide sequence encoding an antibody molecule of theinvention, a heavy or light chain of an antibody, a heavy or light chainvariable domain of an antibody or a portion thereof, or a heavy or lightchain CDR, operably linked to a promoter. Such vectors may include thenucleotide sequence encoding the constant region of the antibodymolecule (see, e.g., PCT Publication WO 86/05807; PCT Publication WO89/01036; and U.S. Pat. No. 5,122,464) and the variable domain of theantibody may be cloned into such a vector for expression of the entireheavy, the entire light chain, or both the entire heavy and lightchains.

[0302] The expression vector is transferred to a host cell byconventional techniques and the transfected cells are then cultured byconventional techniques to produce an antibody of the invention. Thus,the invention includes host cells containing a polynucleotide encodingan antibody of the invention or fragments thereof, or a heavy or lightchain thereof, or portion thereof, or a single chain antibody of theinvention, operably linked to a heterologous promoter. In preferredembodiments for the expression of double-chained antibodies, vectorsencoding both the heavy and light chains may be co-expressed in the hostcell for expression of the entire immunoglobulin molecule, as detailedbelow.

[0303] A variety of host-expression vector systems may be utilized toexpress the antibody molecules of the invention (see, e.g., U.S. Pat.No. 5,807,715). Such host-expression systems represent vehicles by whichthe coding sequences of interest may be produced and subsequentlypurified, but also represent cells which may, when transformed ortransfected with the appropriate nucleotide coding sequences, express anantibody molecule of the invention in situ. These include but are notlimited to microorganisms such as bacteria (e.g., E. coli and B.subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA orcosmid DNA expression vectors containing antibody coding sequences;yeast (e.g., Saccharomyces Pichia) transformed with recombinant yeastexpression vectors containing antibody coding sequences; insect cellsystems infected with recombinant virus expression vectors (e.g.,baculovirus) containing antibody coding sequences; plant cell systemsinfected with recombinant virus expression vectors (e.g., cauliflowermosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed withrecombinant plasmid expression vectors (e.g., Ti plasmid) containingantibody coding sequences; or mammalian cell systems (e.g., COS, 30 CHO,BHK, 293, NSO, and 3T3 cells) harboring recombinant expressionconstructs containing promoters derived from the genome of mammaliancells (e.g., metallothionein promoter) or from mammalian viruses (e.g.,the adenovirus late promoter; the vaccinia virus 7.5K promoter).Preferably, bacterial cells such as Escherichia coli, and morepreferably, eukaryotic cells, especially for the expression of wholerecombinant antibody molecule, are used for the expression of arecombinant antibody molecule. For example, mammalian cells such asChinese hamster ovary cells (CHO), in conjunction with a vector such asthe major intermediate early gene promoter element from humancytomegalovirus is an effective expression system for antibodies(Foecking et al., 1986, Gene 45:101; and Cockett et al., 1990,Bio/Technology 8:2). In a specific embodiment, the expression ofnucleotide sequences encoding antibodies or fragments thereof whichimmunospecifically bind to one or more RSV antigens is regulated by aconstitutive promoter, inducible promoter or tissue specific promoter.

[0304] In bacterial systems, a number of expression vectors may beadvantageously selected depending upon the use intended for the antibodymolecule being expressed. For example, when a large quantity of such aprotein is to be produced, for the generation of pharmaceuticalcompositions of an antibody molecule, vectors which direct theexpression of high levels of fusion protein products that are readilypurified may be desirable. Such vectors include, but are not limited to,the E. coli expression vector pUR278 (Ruther et al., 1983, EMBO12:1791), in which the antibody coding sequence may be ligatedindividually into the vector in frame with the lac Z coding region sothat a fusion protein is produced; pIN vectors (Inouye & Inouye, 1985,Nucleic Acids Res. 13:3101-310^(9;) Van Heeke & Schuster, 1989, J. Biol.Chem. 24:5503-5509); and the like. pGEX vectors may also be used toexpress foreign polypeptides as fusion proteins with glutathione5-transferase (GST). In general, such fusion proteins are soluble andcan easily be purified from lysed cells by adsorption and binding tomatrix glutathione agarose beads followed by elution in the presence offree glutathione. The pGEX vectors are designed to include thrombin orfactor Xa protease cleavage sites so that the cloned target gene productcan be released from the GST moiety.

[0305] In an insect system, Autographa californica nuclear polyhedrosisvirus (AcNPV) is used as a vector to express foreign genes. The virusgrows in Spodoptera frugiperda cells. The antibody coding sequence maybe cloned individually into non-essential regions (for example thepolyhedrin gene) of the virus and placed under control of an AcNPVpromoter (for example the polyhedrin promoter).

[0306] In mammalian host cells, a number of viral-based expressionsystems may be utilized. In cases where an adenovirus is used as anexpression vector, the antibody coding sequence of interest may beligated to an adenovirus transcription/translation control complex,e.g., the late promoter and tripartite leader sequence. This chimericgene may then be inserted in the adenovirus genome by in vitro or invivo recombination. Insertion in a non-essential region of the viralgenome (e.g., region E1 or E3) will result in a recombinant virus thatis viable and capable of expressing the antibody molecule in infectedhosts (e.g., see Logan & Shenk, 1984, Proc. Natl. Acad. Sci. USA 81:355-359). Specific initiation signals may also be required forefficient translation of inserted antibody coding sequences. Thesesignals include the ATG initiation codon and adjacent sequences.Furthermore, the initiation codon must be in phase with the readingframe of the desired coding sequence to ensure translation of the entireinsert. These exogenous translational control signals and initiationcodons can be of a variety of origins, both natural and synthetic. Theefficiency of expression may be enhanced by the inclusion of appropriatetranscription enhancer elements, transcription terminators, etc. (see,e.g., Bittner et al., 1987, Methods in Enzymol. 153:51-544).

[0307] In addition, a host cell strain may be chosen which modulates theexpression of the inserted sequences, or modifies and processes the geneproduct in the specific fashion desired. Such modifications (e.g.,glycosylation) and processing (e.g., cleavage) of protein products maybe important for the function of the protein. Different host cells havecharacteristic and specific mechanisms for the post-translationalprocessing and modification of proteins and gene products. Appropriatecell lines or host systems can be chosen to ensure the correctmodification and processing of the foreign protein expressed. To thisend, eukaryotic host cells which possess the cellular machinery forproper processing of the primary transcript, glycosylation, andphosphorylation of the gene product may be used. Such mammalian hostcells include but are not limited to CHO, VERY, BHK, Hela, COS, MDCK,293, 3T3, W138, BT483, Hs578T, HTB2, BT20 and T47D, NS0 (a murinemyeloma cell line that does not endogenously produce any immunoglobulinchains), CRL7030 and HsS78Bst cells.

[0308] For long-term, high-yield production of recombinant proteins,stable expression is preferred. For example, cell lines which stablyexpress the antibody molecule may be engineered. Rather than usingexpression vectors which contain viral origins of replication, hostcells can be transformed with DNA controlled by appropriate expressioncontrol elements (e.g., promoter, enhancer, sequences, transcriptionterminators, polyadenylation sites, etc.), and a selectable marker.Following the introduction of the foreign DNA, engineered cells may beallowed to grow for 1-2 days in an enriched media, and then are switchedto a selective media. The selectable marker in the recombinant plasmidconfers resistance to the selection and allows cells to stably integratethe plasmid into their chromosomes and grow to form foci which in turncan be cloned and expanded into cell lines. This method mayadvantageously be used to engineer cell lines which express the antibodymolecule. Such engineered cell lines may be particularly useful inscreening and evaluation of compositions that interact directly orindirectly with the antibody molecule.

[0309] A number of selection systems may be used, including but notlimited to, the herpes simplex virus thymidine kinase (Wigler et al.,1977, Cell 11:223), hypoxanthineguanine phosphoribosyltransferase(Szybalska & Szybalski, 1992, Proc. Natl. Acad. Sci. USA 48:202), andadenine phosphoribosyltransferase (Lowy et al., 1980, Cell 22:8-17)genes can be employed in tk-, hgprt- or aprt- cells, respectively. Also,antimetabolite resistance can be used as the basis of selection for thefollowing genes: dhfr, which confers resistance to methotrexate (Wigleret al., 1980, Natl. Acad. Sci. USA 77:357; O'Hare et al., 1981, Proc.Natl. Acad. Sci. USA 78:1527); gpt, which confers resistance tomycophenolic acid (Mulligan & Berg, 1981, Proc. Natl. Acad. Sci. USA78:2072); neo, which confers resistance to the aminoglycoside G-418 (Wuand Wu, 1991, Biotherapy 3:87-95; Tolstoshev, 1993, Ann. Rev. Pharmacol.Toxicol. 32:573-596; Mulligan, 1993, Science 260:926-932; and Morgan andAnderson, 1993, Ann. Rev. Biochem. 62: 191-217; May, 1993, TIB TECH11(5):155-2 15); and hygro, which confers resistance to hygromycin(Santerre et al., 1984, Gene 30:147). Methods commonly known in the artof recombinant DNA technology may be routinely applied to select thedesired recombinant clone, and such methods are described, for example,in Ausubel et al. (eds.), Current Protocols in Molecular Biology, JohnWiley & Sons, NY (1993); Kriegler, Gene Transfer and Expression, ALaboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13,Dracopoli et al. (eds), Current Protocols in Human Genetics, John Wiley& Sons, NY (1994); Colberre-Garapin et al., 1981, J. Mol. Biol. 150:1,which are incorporated by reference herein in their entireties.

[0310] The expression levels of an antibody molecule can be increased byvector amplification (for a review, see Bebbington and Hentschel, Theuse of vectors based on gene amplification for the expression of clonedgenes in mammalian cells in DNA cloning, Vol. 3. (Academic Press, NewYork, 1987)). When a marker in the vector system expressing antibody isamplifiable, increase in the level of inhibitor present in culture ofhost cell will increase the number of copies of the marker gene. Sincethe amplified region is associated with the antibody gene, production ofthe antibody will also increase (Crouse et al., 1983, Mol. Cell. Biol.3:257).

[0311] The host cell may be co-transfected with two expression vectorsof the invention, the first vector encoding a heavy chain derivedpolypeptide and the second vector encoding a light chain derivedpolypeptide. The two vectors may contain identical selectable markerswhich enable equal expression of heavy and light chain polypeptides.Alternatively, a single vector may be used which encodes, and is capableof expressing, both heavy and light chain polypeptides. In suchsituations, the light chain should be placed before the heavy chain toavoid an excess of toxic free heavy chain (Proudfoot, 1986, Nature322:52; and Kohler, 1980, Proc. Natl. Acad. Sci. USA 77:2 197). Thecoding sequences for the heavy and light chains may comprise cDNA orgenomic DNA.

[0312] Once an antibody molecule of the invention has been produced byrecombinant expression, it may be purified by any method known in theart for purification of an immunoglobulin molecule, for example, bychromatography (e.g., ion exchange, affinity, particularly by affinityfor the specific antigen after Protein A, and sizing columnchromatography), centrifugation, differential solubility, or by anyother standard technique for the purification of proteins. Further, theantibodies of the present invention or fragments thereof may be fused toheterologous polypeptide sequences described herein or otherwise knownin the art to facilitate purification.

[0313] 5.7. Kits

[0314] The invention also provides a pharmaceutical pack or kitcomprising one or more containers filled with one or more of theingredients of the pharmaceutical compositions of the invention.Optionally associated with such container(s) can be a notice in the formprescribed by a governmental agency regulating the manufacture, use orsale of pharmaceuticals or biological products, which notice reflectsapproval by the agency of manufacture, use or sale for humanadministration.

[0315] The present invention provides kits that can be used in the abovemethods. In one embodiment, a kit comprises an antibody of theinvention, preferably a purified antibody, in one or more containers. Inan alterative embodiment, a kit comprises an antibody fragment thatimmunospecifically binds to a RSV antigen. In a specific embodiment, thekits of the present invention contain a substantially isolated RSVantigen as a control. Preferably, the kits of the present inventionfurther comprise a control antibody which does not react with the RSVantigen. In another specific embodiment, the kits of the presentinvention contain a means for detecting the binding of an antibody to aRSV antigen (e.g., the antibody may be conjugated to a detectablesubstrate such as a fluorescent compound, an enzymatic substrate, aradioactive compound or a luminescent compound, or a second antibodywhich recognizes the first antibody may be conjugated to a detectablesubstrate). In specific embodiments, the kit may include a recombinantlyproduced or chemically synthesized RSV antigen. The RSV antigen providedin the kit may also be attached to a solid support. In a more specificembodiment the detecting means of the above-described kit includes asolid support to which RSV antigen is attached. Such a kit may alsoinclude a non-attached reporter-labeled anti-human antibody. In thisembodiment, binding of the antibody to the RSV antigen can be detectedby binding of the said reporter-labeled antibody.

[0316] In an additional embodiment, the invention includes a diagnostickit for use in screening serum containing RSV antigens. The diagnostickit includes a substantially isolated antibody specificallyimmunoreactive with a RSV antigen, and means for detecting the bindingof the RSV antigen to the antibody. In one embodiment, the antibody isattached to a solid support. In a specific embodiment, the antibody maybe a monoclonal antibody. The detecting means of the kit may include asecond, labeled monoclonal antibody. Alternatively, or in addition, thedetecting means may include a labeled, competing antigen.

[0317] In one diagnostic configuration, test serum is reacted with asolid phase reagent having a surface-bound RSV antigen obtained by themethods of the present invention. After the RSV antigen binds to aspecific antibody, the unbound serum components are removed by washing,reporter-labeled anti-human antibody is added, unbound anti-humanantibody is removed by washing, and a reagent is reacted withreporter-labeled anti-human antibody to bind reporter to the reagent inproportion to the amount of bound anti-RSV antigen antibody on the solidsupport. Typically, the reporter is an enzyme which is detected byincubating the solid phase in the presence of a suitable fluorometric,luminescent or calorimetric substrate (Sigma, St. Louis, Mo.).

[0318] The solid surface reagent in the above assay is prepared by knowntechniques for attaching protein material to solid support material,such as polymeric beads, dip sticks, 96-well plate or filter material.These attachment methods generally include non-specific adsorption ofthe protein to the support or covalent attachment of the protein,typically through a free amine group, to a chemically reactive group onthe solid support, such as an activated carboxyl, hydroxyl, or aldehydegroup. Alternatively, streptavidin coated plates can be used inconjunction with biotinylated antigen(s).

[0319] Thus, the invention provides an assay system or kit for carryingout this diagnostic method. The kit generally includes a support withsurface-bound recombinant RSV antigen, and a reporter-labeled anti-humanantibody for detecting surface-bound anti-RSV antigen antibody.

6. EXAMPLE KINETIC ANALYSIS OF HUMANIZED RSV MABS BY BIACORE™

[0320] A typical kinetic study involved the injection of 250 μl ofmonoclonal antibody (“Mab”) at varying concentrations (25-300 nM) in PBSbuffer containing 0.05% Tween-20 (PBS/Tween). The flow rate wasmaintained at 75 μl/min, giving a 15 minute dissociation time. Followingthe injection of Mab, the flow was exchanged with PBS/Tween buffer for30 min for determining the rate of dissociation. The sensor chip wasregenerated between cycles with a 1 min pulse of 100 mM HCl. Theregeneration step caused a minimal loss of binding capacity of theimmobilized F-protein (4% loss per cycle). This small decrease did notchange the calculated values of the rate constants for binding anddissociation (also called the k_(on) and k_(off), respectively).

[0321] More specifically, for measurement of k_(assoc) (or k_(on)), Fprotein was directly immobilized by the EDC/NHS method(EDC=N-ethyl-N′-[3-diethylaminopropyl)carbodiimide). Briefly, 25 mg/mlof F protein in 10 mM NaoAc, pH 5.0 was prepared and about a 5-10 μlinjection gives about 30-50 RU (response units) of immobilized F proteinunder the above referenced conditions. The blank was subtracted forkinetic analysis. The column could be regenerated using 100 mM HCl (with60 seconds of contact time being required for full regeneration). Thistreatment removed bound Fab completely without damaging the immobilizedantigen and could be used for over 40 regenerations. For k_(on)measurements, Fab concentrations were 0.39 nM, 0.75 nM, 1.56 nM, 3.13nM, 12.5 nM, 25 nM, 50 nM, and 100 nM. The dissociation phase wasanalyzed for approximately 900 seconds. Kinetics were analyzed by 1:1Langmuir fitting (global fitting). Measurements were done in HBS-EPbuffer (10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% (v/v)Surfactant P20.

[0322] For measurements of combinatorial clones, as disclosed herein,the k_(on) and k_(off) were measured separately. The k_(on) was measuredat conditions that were the same as those for the single mutation clonesand was analyzed similarly.

[0323] For measuring k_(off), the following conditions were employed.Briefly, 4100 RU of F protein were immobilized (as above) withCM-dextran used as the blank. Here, 3000 RU of Fab was bound (withdissociated Fab high enough to offset machine fluctuation). HBS plus 5nM F protein (about 350-2000 times higher than the K_(d)-thedissociation equilibrium constant) was used as buffer. The dissociationphase was 6-15 hours at a flow rate of 5 ml/min. Under the conditionsused herein, re-binding of the dissociated Fab was minimal. For furtherdetails, see the manual with the biosensor.

[0324] The binding of the high affinity anti-RSV antibodies to the Fprotein, or other epitopic sites on RSV, disclosed herein was calculatedfrom the ratio of the first order rate constant for dissociation to thesecond order rate constant for binding or association(K_(d)=k_(off)/k_(on)). The value for k_(on) was calculated based on thefollowing rate equation:

dR/dt=k _(on) [Mab]R _(max)−(k _(on) [Mab]+k _(off))R

[0325] where R and R_(max) are the response units at time t andinfinity, respectively. A plot of dr/dt as a function of R gives a slopeof (k_(a)[Mab]+k_(d))—since these slopes are linearly related to the[Mab], the value k_(on) can be derived from a replot of the slopesversus [Mab]. The slope of the new line is equal to k_(on). Although thevalue of k_(off) can be extrapolated from the Y-intercept, a moreaccurate value was determined by direct measurement of k_(off).Following the injection phase of the Mab, PBS/Tween buffer flows acrossthe sensor chip. From this point, [Mab]=0. The above stated equation fordR/dt thus reduces to:

dr/dt=k or dR/R=k _(off) dt

[0326] Integration of this equation then gives:

ln(R ₀ /R _(t))=k _(off) t

[0327] where R₀/R_(t)) are the response units at time 0 (start ofdissociation phase) and t, respectively. Lastly, plotting In(R₀/R_(t))as a function of t gives a slope of k_(off).

[0328] The numerical values from such antibody variants were as shown inTables 4-7 below. TABLE 4 Summary of Kinetic Constants for High PotencyAntibodies. ANTIBODY K_(on) × 10⁵ (M⁻¹s⁻¹) K_(off) × 10⁻⁴ (s⁻¹) EC₅₀(nM) **SYNAGIS ® 2.04; 1.89; 2.18 7.64; 7.38; 7.02 3.57 **AFFF 1.08;0.96; 1.24 2.74; 2.66; 2.06 *1X-493L1FR 1.85 6.5  *H3-3F4 4.59; 4.67;5.72; 4.45; 4.02 6.25; 5.33 *M3H9 6.05 3.38 *Y10H6 7.57 4.62 *DG 2.65;2.83; 4.16; 1.67; 4.44 3.18; 2.88 *AFFF 2.12; 1.56; 1.86 2.45; 4.46;2.68 *6H8 3.14; 4.44 1.78; 4.73 *L1-7E5 3.29; 3.57; 4.05; 1.92; 3.31;2.29 3.35; 4.26 *L2-15B10 3.69; 2.82; 3.12; 1.34; 4.16; 2.70 5.33; 3.78*P12f2 6.63 2.82 0.65 *P12f4 5.27 2.99 0.70 *P11d4 5.70; 5.72 7.17 >20*Ale9 7.9  4.53 2.5 *A12a6 7.43 2.30 0.62 *A13a11 7.35 2.50 2.04 *A13c47.81; 7.35 2.80 0.52

[0329] TABLE 5 Monoclonal Antibodies vs Bac-F (1:1) Kon (x E + 5) Koff(x E − 5) KD (nM) Chi2 P12f2 4.07 12.8 0.31 (13) 0.9 P12f4 4.95 5.550.11 (35) 0.6 A13c4 3.00 3.96 0.13 (30) 1.2 A12a6 4.60 1.65 0.04 (98)1.2 A1e9 4.33 14.3 0.33 (12) 2.5 A8c7 4.17 8.75 0.21 (19) 1.8 P11d4 4.6628.9 0.62 (6)  1.0 A17d4 4.56 4.07 0.09 (43) 0.5 A4B4 4.34 1.06  0.02(195) 1.5 SYNAGIS ® 1.32 51.5 3.90 (1)  0.6

[0330] TABLE 6 Monoclonal Antibodies vs NUF4 (1:1) Kon (x E + 5) Koff (xE − 5) KD (nM) Chi2 P12f2 5.41 17.8 0.33 (26) 1.2 P12f4 9.43 22.9 0.24(36) 0.9 A13c4 3.65 27.2 0.75 (12) 1.8 A12a6 4.00 29.1 0.73 (12) 1.9A1e9 8.43 58.4 0.69 (13) 0.9 A8c7 8.25 53.5 0.65 (13) 0.7 P11d4 9.0476.6 0.85 (10) 2.5 A17d4 4.99 36.2 0.73 (12) 2.0 A4B4 4.96 28.2 0.57(15) 1.9 SYNAGIS ® 3.04 265 8.70 (1)  0.4

[0331] TABLE 7 Monoclonal Antibodies vs NUF4 (2:1) Kon (x E + 5) Koff (xE − 5) KD (nM) Chi2 P12f2 2.82 23.6 0.84 (371) 1.5 P12f4 2.73 63.6 2.33(134) 4.9 A13c4 3.20 22.5 0.70 (446) 1.7 A12a6 2.18 40.8 1.87 (167) 1.9A1e9 3.29 139 4.22 (74)  2.8 A8c7 4.30 114 2.65 (118) 2.0 P11d4 3.66 3138.55 (36)  3.6 A17d4 2.64 29.2 1.11 (281) 1.7 A4B4 2.03 40.06 2.00 (156)1.4 SYNAGIS ® 0.78 2420  312 (1)  1.3

[0332] 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF, 6H8, L1-7E5, L2-15B10,P12f2, P12f4, P11d4, A1e9, A12a6, A13a11, and A13c4 are Fab fragmentshaving the framework sequences of FIG. 1 and the indicated CDR sequencesindicated listed in Table 2. SYNAGIS AND AFFF are actual monoclonalantibodies with the framework sequences of FIG. 1 and constant regionsas described in Johnson et al. (1997, Journal of Infectious Diseases176:1215-1224) and U.S. Pat. No. 5,824,307. The framework sequences ofthese antibodies may differ slightly from those of the Fab fragments.

[0333] The amino acid sequences of the indicated CDRs in Table 1represent the amino acid residues located at the key locations withinthe CDRs of the high potency antibodies produced by the methodsdescribed herein and in copending applications Ser. Nos. 60/168,426 and60/186,252. For example, to increase the potency of an antibody byproducing a higher k_(on) value, the amino acids located at the keypositions as taught herein by the bold and underlined residues in Table1 for the reference antibody would be replaced by the amino acids listedunder CDRs in Table 2 (and also bold and underlined). Thus, these oneletter codes represent the amino acids replacing the reference aminoacids at the key positions (or critical positions) of the CDRs shown inFIG. 2 (residues in bold in the sequences of Table 2) for a referenceantibody whose potency is to be increased.

7. EXAMPLE MICRONEUTRALIZATION ASSAY

[0334] Neutralization of the antibodies of the present invention weredetermined by microneutralization assay. This microneutralization assayis a modification of the procedures described by Anderson et al. (1985,J. Clin. Microbiol. 22:1050-1052, the disclosure of which is herebyincorporated by reference in its entirety). The procedure used here isdescribed in Johnson et al., 1999, J. Infectious Diseases 180:35-40, thedisclosure of which is hereby incorporated by reference in its entirety.Antibody dilutions were made in triplicate using a 96-well plate. TenTCID₅₀ of respiratory syncytial virus (RSV-Long strain) were incubatedwith serial dilutions of the antibody (or Fabs) to be tested for 2 hoursat 37° C. in the wells of a 96-well plate. RSV susceptible HEp-2 cells(2.5×10⁴) were then added to each well and cultured for 5 days at 37° C.in 5% CO₂. After 5 days, the medium was aspirated and cells were washedand fixed to the plates with 80% methanol and 20% PBS. RSV replicationwas then determined by F protein expression. Fixed cells were incubatedwith a biotin-conjugated anti-F protein monoclonal antibody (pan Fprotein, C-site-specific MAb 133-1H) washed and horseradish peroxidaseconjugated avidin was added to the wells. The wells were washed againand turnover of substrate TMB (thionitrobenzoic acid) was measured at450 nm. The neutralizing titer was expressed as the antibodyconcentration that caused at least 50% reduction in absorbency at 450 nm(the OD₄₅₀) from virus-only control cells. The results from the assayfor the monoclonal antibodies and Fab fragments listed in Table 2 areshown in Table 4 supra and Table 8 infra. TABLE 8 End Point RSVMicroneutratization Titer Of High On Rate Mutant IgG and Fab Fold FoldMean Differ- Mean Differ- IC50 STDEV ence IC50 STDEV ence n (Curve)Curve (Curve (Control) Control (Control (assay Molecule μg/ml IC50ICX50) μg/ml IC50 IC50) repeat) **SYNAGIS ® 0.4527 0.208 — 0.5351 0.238— 8 **A1e9 0.0625 0.0268 7 0.0645 0.0223 8 3 **A17d4 0.0342 0.022 130.0354 0.0187 15 4 **P11d4 0.0217 0.0331 21 0.0289 0.0110 19 5 **P12f20.0231 0.0141 20 0.0223 0.0083 24 6 **A8c7 0.0337 0.0309 13 0.03830.0283 14 5 **A12a6 0.0357 0.0316 13 0.0354 0.0261 15 7 **P12f4 0.02420.0163 19 0.0235 0.0076 23 7 **A13c4 0.0376 0.0268 12 0.0375 0.0213 14 6**A4B4 0.0171 0.0018 27 0.0154 0.00417 35 2 *A1e9 0.157 — 3 0.125 — 4 1*A17d4 0.0179 — 25 0.0171 — 31 1 *P11d4 >1.00 — — >1.00 — — 1 *P12f20.0407 0.0112 11 0.0326 0.00905 16 2 *A8c7 0.177 — 3 0.157 — 34 1 *A12a60.0287 0.00417 16 0.0310 0.00982 17 2 *P12f4 0.0464 0.00791 10 0.03510.0126 15 2 *A13c4 0.0264 0.00141 17 0.0258 0.00071 21 2 *A4B4 0.0414 —11 0.0411 — 13 1 *A13a11 0.120 0.0222 4 0.1022 0.0260 5 2 *A1h5 0.1940.462 2 0.176 0.0625 3 2

8. EXAMPLE RSV FUSION INHIBITION ASSAY

[0335] The ability of the antibodies of the invention or fragmentsthereof to block RSV-induced fusion after viral attachment to the cellsis determined in a fusion inhibition assay. This assay is identical tothe microneutralization assay, except that the cells were infected withRSV (Long) for four hours prior to addition of antibody (Taylor etal,1992, J. Gen. Virol. 73:2217-2223).

9. EXAMPLE ISOTHERMAL TITRATION CALORIMETRY

[0336] Thermodynamic binding affinities and enthalpies were determinedfrom isothermal titration calorimetry (ITC) measurements on theinteraction of antibodies with RSV F glycoprotein (NUF4), an antigenwhich mimics the binding site of the RSV virus.

Methods & Materials

[0337] Antibodies & Antigen A13c4, A17d4, A4B4, and SYNAGIS® werediluted in dialysate and the concentrations were determined by UVspectroscopic absorption measurements with a Perkin-Elmer Lambda 4BSpectrophotometer using an extinction coefficient of 217,000 M⁻¹ cm⁻¹ atthe peak maximum at 280 nm. The diluted NUF4 concentrations werecalculated from the ratio of the mass of the original sample to that ofthe diluted sample since its extinction coefficient was too low todetermine an accurate concentration without employing and losing a largeamount of sample.

[0338] ITC Measurements

[0339] The binding thermodynamics of the antibodies were determined fromITC measurements using a Microcal, Inc. VP Titration Calorimeter. The VPtitration calorimeter consists of a matched pair of sample and referencevessels (1.409 ml) enclosed in an adiabatic enclosure and a rotatingstirrer-syringe for titrating ligand solutions into the sample vessel.The ITC measurements were performed at 25° C. and 35° C. The samplevessel contained the antibody in the phosphate buffer while thereference vessel contained just the buffer solution. The phosphatebuffer solution was saline 67 mM PO₄ at pH 7.4 from HyClone, Inc. Fiveor ten μl aliquots of the 0.05 to 0.1 mM NUF4 solution were titrated 3to 4 minutes apart into the antibody sample solution until the bindingwas saturated as evident by the lack of a heat exchange signal. Withsome antibody sample solutions, additional constant amounts of heat withthe addition of each aliquot were observed following binding saturationof the antibody. This was attributed to a heat of dilution of the NUF4titrant and was subtracted from the titrant heats obtained during thetitration prior to analysis of the data.

[0340] A non-linear, least square minimization software program fromMicrocal, Inc., Origin 5.0, was used to fit the incremental heat of theith titration (ΔQ (i)) of the total heat, Q_(t), to the total titrantconcentration, X_(t), according to the following equations (I),

Q _(t) =nC _(t) ΔH _(b) °V{1+X _(t) /nC _(t)+1/nK _(b) C _(t)−[(1+X _(t)/nC _(t)+1/nK _(b) C _(t))²−4X _(t) /C _(t]) ^(½)}/2   (1a)

ΔQ(i)=Q(i)+dVi/2V {Q(i)+Q(i−1)}−Q(i−1)   (1b)

[0341] where C_(t) is the initial antibody concentration in the samplevessel, V is the volume of the sample vessel, and n is the stoichiometryof the binding reaction, to yield values of K_(b), ΔH_(b)°, and n. Theoptimum range of sample concentrations for the determination of K_(b)depends on the value of K_(b) and is defined by the followingrelationship.

C_(t)K_(b)n≦500   (2)

[0342] so that at 1 μM the maximum K_(b) that can be determined is lessthan 2.5×10⁸ M⁻¹. If the first titrant addition did not fit the bindingisotherm, it was neglected in the final analysis since it may reflectrelease of an air bubble at the syringe opening-solution interface.

Results

[0343] The ITC results are summarized in Table 9. The higher than 2stoichiometries in Table 9 indicate that either the concentrationdetermination of the antibody or NUF4 was incorrect. Since the same NUF4sample was used as a titrant with antibodies having the amino acidsequence of A13c4 at 35° C. and A17d4 at 35° C., which exhibit in atleast one of the titrations the correct stoichiometry of 2, it isassumed that the titrant concentration was correct and that the largevalues of n result from incorrectly determined antibody concentrations.However, it can be shown that the binding constants are criticallydependent on the titrant concentration and, thus, despite the 2-3disparity in n, the binding constants are correct. Since the bindingconstants of antibodies having the amino acid sequence of A4B4 and A13c4at 25° C. were near the upper determination limit by ITC (equation 2)and with the limited amount of available NUF4, it was decided to use 35°C. as the reference temperature for comprising the binding affinities.The results summarized in Table 9 show that the binding affinities toNUF4 are in the order A4B4>A13c4>A17d4> SYNAGIS®. TABLE 9 AverageBinding Constants and Enthalpies of NUF4 to Antibodies Antibody K_(b)ΔH_(b) in kJ mol⁻¹ A4B4 269 ± 74 × 10⁶ M⁻¹ or ˜3.7 nM* 92.8 ± 1.0  A13c4107 ± 28 × 10⁶ M⁻¹ or 9 nM 67 ± 17 A17d4 75 ± 14 × 10⁶ M⁻¹ or 13 nM 68 ±10 SYNAGIS ® 1.23 ± 0.17 × 10⁶ M⁻¹ or 810 nM 71 ± 5 

10. EXAMPLE COTTON RAT PROPHYLAXIS

[0344] To determine the ability of SYNAGIS® to prevent lower respiratorytract RSV infection in cotton rats when administered by and intravenous(IV) route and to correlate the serum concentration of SYNAGIS® with areduction in lung RSV titer.

Materials & Methods

[0345] SYNAGIS® lot L94H048 was used for studies III-47 and III-47A.SYNAGIS® lot L95 K016 was used for study III-58. Bovine serum albumin (BSA) (fraction V, Sigma Chemicals). RSV-Long (A subtype) was propagatedin Hep-2 cells.

[0346] On day 0, to groups of cotton rats (Sigmodon hispidis, averageweight 100 g) were administered SYNAGIS®, RSV-IGIV or BSA wasadministered by intramuscular injection. Twenty-four hours postadministration, the animals were bled and infected intranasally with 10⁵pfu of RSV. Twenty-four hours later, the animals were bled and infectedintranasally with 10⁵ PFU or RSV (Long Strain). Four days after theinfection, animals were sacrificed, I10 and their lung tissue washarvested and pulmonary virus titers were determined by plaquetitration. For studies III-47 and III-47A, the doses of monoclonalantibody (“MAb”) consisted of 0.31, 0.63, 1.25, 2.5, 5.5 and 10 mg/kg(body weight). For studies III-58, the doses of MAb consisted of 0.63,1.25, 2.5, 5.5 and 10 mg/kg (body weight). In all three studies bovineserum albumin (BSA) 10 mg/kg was used as a negative control. Humanantibody concentrations in the serum at the time of challenge aredetermined using a sandwich ELISA.

Results

[0347] The results of the individual experiments are presented in Tables10-12. The results of all of the experiments combined is shown in Table13. All three studies show a significant reduction of pulmonary virustiters in animals treated with SYNAGIS®. A clear dose-response effectwas observed in the animals. The combined data indicated that a dose of2.5 mg/kg results in a greater than 99% reduction in lung RSV titer. Themean serum concentration of SYNAGIS® for this dose at the time of viralchallenge was 28.6 μg/ml. TABLE 10 EXPERIMENT III-47 Mean ± Std ErrorLung Viral Titer Number of Concentration of Geometric Mean ± StdCompound Animals Dose Human IgG (μg/ml) Error (log10 pfu/gm) BSA 4 0 1.4× 10⁵ ± 1.7 SYNAGIS ® 3 0.312 mg/kg 3.83 ± 1.1  2.1 × 10⁴ ± 2.1SYNAGIS ® 3 0.625 mg/kg 5.27 ± 0.37 7.7 × 10⁴ ± 1.6 SYNAGIS ® 4  1.25mg/kg 9.15 ± 0.16 3.4 × 10⁴ ± 1.3 SYNAGIS ® 3  2.50 mg/kg 23.4 ± 2.8 1.4 × 10³ ± 1.7 SYNAGIS ® 2  5.0 mg/kg 42.4 ± 13.4 4.6 × 10² ± 4.6SYNAGIS ® 4  10.0 mg/kg 141.1 ± 14.4  1.0 × 10² ± 1.0

[0348] TABLE 11 EXPERIMENT III-47A Mean ± Std Error Lung Viral TiterNumber of Concentration of Geometric Mean ± Std Compound Animals DoseHuman IgG (μg/ml) Error (log10 pfu/gm) BSA 4 0 1.9 × 10⁵ ± 1.2 SYNAGIS ®4 0.312 mg/kg  1.8 ± 0.12 8.5 × 10⁴ ± 1.2 SYNAGIS ® 4 0.625 mg/kg  4.0 ±0.19 5.0 × 10⁴ ± 1.6 SYNAGIS ® 4  1.25 mg/kg 11.8 ± 0.68 1.9 × 10³ ± 1.4SYNAGIS ® 4  2.50 mg/kg 18.9 ± 2.0  5.3 × 10³ ± 1.6 SYNAGIS ® 3  5.0mg/kg 55.6 ± 2.3  1.6 × 10² ± 1.3 SYNAGIS ® 4  10.0 mg/kg 109.7 ± 5.22 1.0 × 10² ± 1.0

[0349] TABLE 12 EXPERIMENT III-58 Mean ± Std Error Lung Viral TiterNumber of Concentration of Geometric Mean ± Std Compound Animals DoseHuman IgG (μg/ml) Error (log10 pfu/gm) BSA 4 0 1.1 × 10⁵ ± 1.2 SYNAGIS ®4 0.625 mg/kg 5.78 ± 0.32 1.6 × 10⁴ ± 1.2 SYNAGIS ® 4  1.25 mg/kg 9.82 ±0.23 1.6 × 10³ ± 1.3 SYNAGIS ® 4  2.50 mg/kg 34.1 ± 2.11 4.3 × 10² ± 1.6SYNAGIS ® 3  5.0 mg/kg 58.3 ± 4.48 1.0 × 10² ± 1.0 SYNAGIS ® 4  10.0mg/kg 111.5 ± 5.04  1.0 × 10² ± 1.0

[0350] TABLE 13 EXPERIMENT III-47, III-47A and III-58 COMBINED Mean ±Std Error Lung Viral Titer Number of Concentration of Geometric Mean ±Std Compound Animals Dose Human IgG (μg/ml) Error (log10 pfu/gm) BSA 180 1.3 × 10⁵ ± 1.2 SYNAGIS ® 7 0.312 mg/kg 2.67 ± 0.60 4.6 × 10⁴ ± 1.5SYNAGIS ® 17 0.625 mg/kg 5.27 ± 0.27 2.7 × 10⁴ ± 1.3 SYNAGIS ® 18  1.25mg/kg 10.1 ± 0.29 3.3 × 10³ ± 1.4 SYNAGIS ® 17  2.50 mg/kg 28.6 ± 2.159.6 × 10² ± 1.5 SYNAGIS ® 15  5.0 mg/kg 55.6 ± 3.43 1.3 × 10² ± 1.2SYNAGIS ® 18  10.0 mg/kg 117.6 ± 5.09  1.0 × 10² ± 1.0

11. EXAMPLE INTRAMUSCULAR COTTON RAT STUDIES

[0351] This experiment demonstrates that a greater reduction in RSVtiter is achieved when A4b4, A4b4-F52S or A4b4/L1FR-S28R is administeredintramuscularly to a cotton rat than when the same concentration ofSYNAGIS® is administered intramuscularly to a cotton rat.

Materials & Methods

[0352] Intramuscular Cotton Rat Prophylaxis

[0353] Cotton rats (S. hispidus, average weight 100 grams) wereanesthetized with methoxyflurane and dosed with 0.1 ml of purifiedmonoclonal antibody (MAb) or BSA control by intramuscular (i.m.)injection. Twenty-four hours later animals were again anesthetized, bledfor serum MAb concentration determination, and challenged with 10⁵ PFURSV long by intranasal (i.n.) instillation. Four days later animals weresacrificed, serum samples were obtained, and their lungs were harvested.Lungs were homogenized in 10 parts (wt/vol) of Hanks Balanced Saltsolution and the resultant suspension was used to determine pulmonaryviral titers by plaque assay.

[0354] Intramuscular Cotton Rat Pharmacokinetics

[0355] Cotton rats (S. hispidus, average weight 100 grams) wereanesthetized with methoxyflurane and dosed with 0.1 ml of purified MAbor BSA control by intramuscular (i.m) injection. Twenty-four hours laterall of the animals were bled for serum MAb concentration determination,and half of the animals from each group were sacrificed to performbronchoalveolar lavage (BAL). Four days later the remaining animals weresacrificed, serum samples were obtained and BAL performed.

Results

[0356] As shown in Tables 14-16, a greater reduction in RSV titer isachieved with equivalent or lower lung levels of A4b4, A4b4-F52S, orA4b4/L1FR-S28R as with SYNAGIS®. TABLE 14 Intramuscular Cotton RatProphylaxis Data 0.5 mg/kg 0.125 mg/kg Serum Lung Virus Iog Serum LungVirus Iog IgG IgG Titer Virus IgG IgG Titer Virus (μg/ml) (μg/ml)(pfu/gm) Titer (μg/ml) (μg/ml) (pfu/gm) Titer Synagis 3.4 0.099 7.3 ×10³ 3.9 0.893 0.024 3.1 × 10⁴ 4.5 A4b4-F52S 2.9 0.089 7.3 × 10² 2.90.781 0.020 8.6 × 10³ 3.9 A4b4/L1FR-S28R 3.3 0.093 6.1 × 10² 2.8 0.7480.016 2.3 × 10⁴ 4.4 BSA 5.9 × 10⁴ 4.8

[0357] TABLE 15 Intramuscular Cotton Rat Prophylaxis Data 0.5 mg/kg 1mg/kg Serum Lung Iog (10) Serum Lung Iog (10) IgG IgG Lung IgG IgG LungMolecule (μg/ml) (μg/ml) Virus (μg/ml) (μg/ml) Virus A4b4 2.4 0.013 4.33.1 0.094 3.4 Synagis 1.9 0.038 4.4 4.2 0.212 3.3 BSA 4.4

[0358] TABLE 16 Intramuscular Cotton Rat Pharmacokinetics Data 24 Hours96 hours Serum IgG BAL IgG Serum lgG BAL IgG Molecule (μg/ml) (ng/ml)(μg/ml) (ng/ml) A4b4 3.4 2.2 2.6 1.4 Synagis 4.1 5.3 2.8 3.5

12. EXAMPLE IN VITRO ISOLATION OF SYNAGIS® SPECIFIC MONOCLONAL ANTIBODYRESISTANT MUTANTS(MARM) OF RSV STRAIN A/LONG

[0359] This example demonstrates that MARMs can be isolated from RSVlaboratory strain A/Long upon multiple rounds of selection in thepresence of SYNAGIS®.

Materials

[0360] SYNAGIS® (formulated product: ref RN1002.148), Control Pan RSVMAb Pool (Chemicon MAB858-4, a blend of three MAbs anti-F, G, and Nproteins), anti-RSV Type A MAb (Chemicon MAB858-1), and anti-RSV Type BMAb (Chemicon MAB858-2) were used in this study. Secondary detectingreagents were either Alexa™488 conjugated Goat F(ab′)₂ anti- Mouse orHuman IgG (H+L). RSV A/Long strain (5×10⁷ TCID₅/ml) was propagated fromMaster Virus Bank 031797. HEp-2 cells were propagated in EMEMsupplemented with 10% FBS and 2 mM L-Gln in a 37° C., 5% CO₂environment. Total cellular RNAs were isolated from infected HEp-2 cellswith the Promega RNAgents Kit. cDNAs were synthesized with theBoehringer Mannheim 1^(st) Strand cDNA Synthesis Kit, using Oligo- (dT)primer. Amplification of a fragment of the F protein for DNA sequencingwas performed by Polymerase Chain Reaction (PCR), using gene-specificoligonucleotides (MR-120 and MR-1 22) and the Boehringer Mannheim HighFidelity PCR Kit.

Methods

[0361] Selection of MARMs:

[0362] 4×10⁵ HEp-2 cells were seeded per well of a 24-well plate ingrowth medium (EMEM, 10% FBS, 2 mM L-Gln) and incubated overnight ingrowth conditions (37° C., 5% CO₂). 40 wells were seeded for individualMARM selection. Prior to infection, a fresh vial of master virus bankRSV A/Long was thawed rapidly at 37° C. and the virus titer adjusted to4×10⁶ pfu/ml in HEp-2 growth medium. SYNAGIS® was added to the virusinoculum at a final concentration of 30 μg/ml, and the mixture wasincubated at 37° C. for one hour. An aliquot of RSV was incubated withan irrelevant Human IgG1 Mab (MEDI-507) and was used as a negativecontrol. An uninfected control well was also set up for each plate.Cells were washed once with fresh medium and were overlayed with 100 μlof RSV A/Long virus stock/MAb mixture (multiplicity of infection[m.o.i.]=4). Cells were incubated for four hours in growth conditionsfollowed by addition of 1 ml of growth medium to each well. Cytopathiceffect (CPE) was monitored on a daily basis by light microscopy.Following seven days of selection the contents of each well weresupplemented with a further 30 μg/ml of SYNAGIS® or irrelevant HumanIgG1 Mab, and were used to infect freshly seeded HEp-2 cells (4×10⁵cells/ well of 24-well plate). After a further seven days of selectionthe process was repeated one more time for a total of three rounds ofselection.

[0363] Plaque Purification of MARMs:

[0364] After the third round of selection, the contents of tenindependent wells were chosen at random and used for plaque purificationof MARMs. The remaining supernatants were mixed 1:1 with 50% sucrosesolution and immediately frozen at −80° C. Supernatants were diluted1:10, 1:100, and 1:1000 in growth medium and incubated with 30 μg/ml ofSYNAGIS® for one hour prior to infection. 1 ml of virus inoculum wasoverlayed on monolayers of HEp-2 cells in 60 mm round culture dishes(1×10⁶ cells/dish) and incubated for 4 hours in growth conditions.Following infection the inoculum was carefully aspirated and cells wereoverlayed with 3 ml growth medium supplemented with 0.8% low meltingtemperature agarose (Gibco BRL), and containing 30 μg/ml of SYNAGIS®.Dishes were returned to the incubator after complete solidification ofagarose and monitored daily for plaque formation. Uninfected controlsand wild type RSV control plates were set up for comparison. After fiveto six days, each plate was overlayed with an additional 2 ml of 0.8%agarose in growth medium supplemented with 50 μ/ml neutral red.Following an overnight incubation in growth conditions, plaques werescored and picked for a second round of purification.

[0365] Amplification of clonal MARMs:

[0366] Following a second round of plaque purification two clones fromeach isolate were expanded for production of high titer viral stocks.Individual plaques were picked with the aid of a pipet tip and incubatedin 0.1 ml of fresh medium at 4° C. overnight to elute the virus. Eachaliquot was used to infect HEp-2 cells in flat bottom 96-well plates(1×10⁴ cells/well) in the presence of 30 μg/ml SYNAGIS®. After five daysthe entire well content was used to infect HEp-2 cells in the presenceof 30 μg/ml of SYNAGIS® in 24-well plates. Each inoculum wassubsequently expanded in the presence of 30 μ/ml of SYNAGIS® to a T-25flask (1×10⁶ cells/ flask) and 5 mls of high titer virus stock wereproduced. MARM stocks were frozen as described above.

[0367] F Protein cDNA Synthesis and DNA Sequencing:

[0368] In order to determine the nucleotide sequence of an approximately400 nucleotide region of the F protein gene thought to contain theepitope for SYNAGIS®, First strand cDNA synthesis was performed withtotal cellular RNAs isolated from MARM-infected HEp-2 cells at 4 dayspost-infection. Amplification of a fragment of the F protein for DNAsequencing was performed as described in Materials section. AmplifiedMARM F-protein cDNAs were purified by phenol/chloroform extraction andethanol precipitation, and used in a PCR sequencing reaction withgene-specific oligos and Perkin-Elmer Cetus Big-Dye Terminator reactionmix to sequence the relevant region.

[0369] Immunofluorescence Assay (IFA:

[0370] Cells infected with RSV isolates were tested for anti-RSV bindingby SYNAGIS® and Control Pan RSV MAb Pool, as follows. Four to five dayRSV-infected HEp-2 cultures were mixed with PBS and centrifuged at 300×gfor 5 minutes at room temperature. Pellets were resuspended in a smallvolume of PBS for analysis. Five to ten microliters of each cellsuspension were spotted per 5 mm well on acetone washed 12-well HTCsuper cured glass slides and allowed to air dry. Slides were fixed incold (−20° C.) acetone for 10 minutes. Reactions were blocked byspotting 10 μl of 1% BSA in PBS into each well and incubating for 10minutes at room temperature. Slides were washed three times in1×PBS/0.1% Tween-20 and air-dried. Ten microliters of each primaryantibody reagent diluted to 250 ng/ml in blocking buffer were spottedper well and reactions were incubated in a humidified 37° C. environmentfor 30 minutes. Slides were then washed for 1 minute in three changes of1×PBS/0.1% Tween-20 and were air-dried. Ten microliters of appropriatesecondary conjugated antibody reagent diluted to 250 ng/ml in blockingbuffer were added to each respective well and reactions were incubatedin a humidified 37° C. environment for an additional 30 minutes. Slideswere then washed for 1 minute in three changes of 1×PBS/0.1% Tween-20.Five micro liters of 50% glycerol in PBS, 10 mM Tris, pH 8.0, 1 mM EDTAwere spotted in each reaction well, and slides were mounted with coverslips. Each reaction well was subsequently analyzed by fluorescencemicroscopy at 200×power using a B-2A filter (EX 450-490 nm). Positivereactions were scored against an auto-fluorescent background obtainedwith unstained cells or cells stained with secondary reagent alone.

[0371] RSV positive reactions were characterized by bright fluorescencepunctuated with small inclusions in the cytoplasm of infected cells.

[0372] Microneutralization Assay:

[0373] The procedure used here is described in Johnson et al., 1999, J.Infectious Diseases 180:35-40, the disclosure of which is herebyincorporated by reference in its entirety. Briefly, antibody dilutionswere made in triplicate using a 96-well plate. Ten TCID₅₀ of RSV A MARMSwere incubated with serial dilutions of the antibody to be tested for 2hours at 37° C. in the wells of a 96-well plate. The antibodies used inthe assay comprised the heavy chain of SYNAGIS®, the heavy chain ofSYNAGIS® with a point mutation or the heavy chain of A4B4, and the lightchain of SYNAGIS®, the light chain of SYNAGIS®with point mutations, thelight chain of A4B4 with point mutations, the light chain of A4B4 withpoint mutations, the light chain of L1FR (a.k.a. 1X-493L1FR) or thelight chain of L1FR with point mutations. Mab 13/19 was used in theassay as a positive control. RSV susceptible HEp-2 cells (2.5×10⁴) werethen added to each well and cultured for 5 days at 37° C. in 5% CO₂.After 5 days, the medium was aspirated and cells were washed and fixedto the plates with 80% methanol and 20% PBS. RSV replication was thendetermined by F protein expression. Fixed cells were incubated with abiotin-conjugated anti-F protein monoclonal antibody (pan F protein,C-site-specific MAb 133-1H) washed and horseradish peroxidase conjugatedavidin was added to the wells. The wells were washed again and turnoverof substrate TMB (thionitrobenzoic acid) was measured at 450 nm. Theresults from the assay for the monoclonal antibodies are shown in Table19 infra.

Results and Discussion

[0374] The binding activity of SYNAGIS® was tested by IFA against apanel of 20 RSV A/Long MARMs obtained by three rounds of selection onHEp-2 cells in the presence of SYNAGIS®. A pool of monoclonal antibodiesagainst the fusion, glycoprotein, and nuclear proteins of RSV (ControlPan RSV MAb Pool) was used as a positive control for detection of RSV.Subtyping of RSV MARMs was performed with two monoclonal antibodies thatdistinguish type A and type B glycoproteins. As summarized in Table 17,lack of binding activity by SYNAGIS® was demonstrated in all 20 MARMs.Contrasting with lack of binding by SYNAGIS®, binding of the Control PanRSV MAb Pool was demonstrated for all MARMs tested. All 20 MARMs wereclassified as RSV type A. Wild type RSV A/Long infected HEp-2 cellsbound SYNAGIS®, Pan RSV MAb Pool, and RSV Type A MAb, but failed toreact with RSV Type B MAb, as expected.

[0375] DNA sequencing analysis of an approximately 400-nucleotide regionof the RSV F protein cDNA encompassing the proposed SYNAGIS® epitoperevealed a single mutation at the amino-acid level at position 272.Table 18 shows the amino acid change at position 272 in twelve isolatessequenced to date. Although the entire nucleotide sequence of RSV MARMsF protein has not been determined, these results suggest that amino acid272 is a critical residue in modulating the binding of SYNAGIS® to itsepitope.

[0376] The ability of various monoclonal antibodies to neutralize thereplication of RSV A MARMs was determined. As shown in Table 19, theability of the monoclonal antibodies to neutralize the replication ofRSV MARMs varied depending upon the amino acid sequence of the heavychain (HC) and light chain (LC) of the antibody. TABLE 17Characterization of anti-RSV Binding Activity by SYNAGIS ®, Control PanRSV MAb Pool (anti-F, G, N proteins), anti-RSV Type A MAb, and anti-RSVType B MAb by Immunofluorescence Assay (IFA) on RSV A/Long MARMs.Reactivity w/ Reactivity w/ Reactivity w/ Reactivity w/ anti-RSVanti-RSV Anti-RSV RSV MARM SYNAGIS ® MAb Pool type A MAb type B MAb B1 − + + − B2  − + + − B3  − + + − B4  − + + − B5  − + + − B6  − + + − B7 − + + − B8  − + + − B9  − + + − B10 − + + − B11 − + + − B12 − + + − B13− + + − B14 − + + − B15 − + + − B16 − + + − B17 − + + − B18 − + + − B19− + + − B20 − + + − Wt RSV A/ + + + −

[0377] TABLE 18 Amino acid sequence of a region of the wild type RSVA/Long and MARMs F Protein encompassing the proposed SYNAGIS ® epitope.Isolate Amino Acid Sequence (Position number) Wild type 266 267 268 269270 271 272 273 274 275 276 277 278 279 RSV Long I T N D Q K K L M S N NV Q MARM I T N D Q K N L M S N N V Q B1 MARM I T N D Q K M L M S N N V QB2 MARM I T N D Q K M L M S N N V Q B3 MARM I T N D Q K M L M S N N V QB4 MARM I T N D Q K M L M S N N V Q B6 MARM I T N D Q K T L M S N N V QB7 MARM I T N D Q K M L M S N N V Q B8 MARM I T N D Q K Q L M S N N V QB9 MARM I T N D Q K T L M S N N V Q B10 MARM I T N D Q K M L M S N N V QB13 MARM I T N D Q K Q L M S N N V Q B14 MARM I T N D Q K M L M S N N VQ B15

[0378] TABLE 19 MARM analysis by Microneutralization Assay RSVMicroneutralization Assay on RSV A MARMs to Synagis with RSV MonoclonalAntibody Amino Acid Change K272Q K272N K272M K272T Antibody MARMs toSYNAGIS ® HC/LC MARM B9 MARM B1 MARM B2 MARM B7 Note SYNAGIS ®/SYNAGIS ®− − − − Purified Ab A4B4/A4B4 + + + + Purified Ab A4B4/SYNAGIS ® + + + −Purified Ab SYNAGIS ®/A4B4 − − − − Purified Ab A4B4/L1FR + + + −Purified Ab A4B4/L1FR S28R + + + + Purified Ab A4B4/L1FR S28R,S52F + + + + Purified Ab A4B4/L1FR-28R, 52F, F93 − − − − Purified AbA4B4/A4b428S + + + + Purified Ab A4B4/A4b452S + + + + Purified Ab Mab13/19 + + + + Purified Ab SYNAGIS ®/K53F − − − − 293H sup SYNAGIS ®/S28R− − − − 293H sup SYNAGIS ®/Q26S − − − − 293H sup SYNAGIS ®/A55D − − − −293H sup SYNAGIS ®/K24S − − − − 293H sup SYNAGIS ®/C25A − − − − 293H supSYNAGIS ®/L27S − − − − 293H sup SYNAGIS ®/S52F − − − − 293H supSYNAGIS ®/L105V − − − − 293H sup T98F/SYNAGIS ® − − − − Purified abS32A/SYNAGIS ® − − − − 293H sup S95D/SYNAGIS ® − − − − 293H supD58H/SYNAGIS ® − − − − 293H sup A105Q/SYNAGIS ® − − − − 293H supS65D/SYNAGIS ® − − − − 293H sup W100F/SYNAGIS ® − − − − 293H sup

13. EXAMPLE IN VITRO ISOLATION OF A4B4 SPECIFIC MONOCLONAL ANTIBODYRESISTANT MUTANTS (MARM) OF RSV STRAIN A/LONG

[0379] This example demonstrates that MARMs can be isolated from RSVlaboratory strain A/Long upon multiple rounds of selection in thepresence of A4B4

Materials

[0380] A4B4 (Lot #524-9, 3.57 mg /ml), Control Pan RSV MAb Pool(Chemicon MAB858-4, a blend of three MAbs anti-F, G, and N proteins),anti-RSV Type A MAb (Chemicon MAB858-1), and anti-RSV Type B MAb(Chemicon MAB858-2) were used in this study. Secondary detectingreagents were either Alex™488 conjugated Goat F (ab′)₂ anti-Mouse orHuman IgG (H+L). Virus bank of RSV A/Long strain NWVB020500 (2.38×10⁷TCID₅₀/ml) propagated from Master Virus Bank 031797. HEp-2 cells werepropagated in EMEM supplemented with 10% FBS and 2 mM L-Gln in a 37° C.,5% CO₂ environment. Messanger RNA was purified from the infected cellsusing the mRNA Capture Kit (Roche). The mRNA samples were used to makecDNA using the reagents from the cDNA First Strand Reacition Kit(Roche), followed by amplification of the RSV F protein gene by thePolymerase Chain Reaction (PCR) using gene specific primers.

Methods

[0381] Selection of MARMs:

[0382] 4×10⁵ HEp-2 cells were seeded per well of a 24-well plate ingrowth medium (EMEM, 10% FBS, 2 mM L-Gln) and incubated overnight ingrowth conditions (37° C., 5% CO₂). 44 wells were seeded for individualMARM selection. Prior to infection, a fresh vial of virus bank RSVA/Long was thawed rapidly at 37° C. and the virus titer adjusted to4×10⁶ pfu/ml in HEp-2 growth medium. A4B4 was added to the virusinoculum at a final concentration of 2 μg/ml, and the mixture wasincubated at 37° C. for one hour. An aliquot of RSV was incubated withan irrelevant Human IgG1 Mab (MEDI-507) and was used as a negativecontrol. An uninfected control well was also set up for each plate.Cells were washed once with fresh medium and were overlayed with 100 μlof RSV A/Long virus stock/MAb mixture. Cells were incubated for fourhours in growth conditions followed by addition of 1 ml of growth mediumto each well. Cytopathic effect (CPE) was monitored on a daily basis bylight microscopy. Following seven days of selection the contents of eachwell were supplemented with a further 4 μg/ml of A4B4 or irrelevantHuman IgG1 Mab, and were used to infect freshly seeded HEp-2 cells(4×10⁵ cells/ well of 24-well plate). After a further seven days ofselection the process was repeated one more time for a total of threerounds of selection. The contents of wells showing clear CPE were mixed1:1 with 50% sucrose solution and immediately frozen at −80° C.

[0383] Plaque Purification of MARMs:

[0384] After the third round of selection, the contents of twoindependent wells were chosen at random and used for plaque purificationof MARMs. A fresh vial of MARM stock (was frozen after third round ofselection) was thawed at room temperature and was diluted 1:10, 1:100,and 1:1000 in growth medium and incubated with 4 μg/ml of A4B4 for onehour prior to infection. 0.5 ml of virus inoculum was overlayed onmonolayers of HEp-2 cells in 6 well plates (5×10⁵ cells/ well) andincubated for 4 hours in growth conditions. Following infection theinoculum was carefully aspirated and cells were overlayed with 2 mlgrowth medium supplemented with 0.8% low melting temperature agarose(Gibco BRL), and containing 4 μg/ml of A4B4. Dishes were returned to theincubator after complete solidification of agarose and monitored dailyfor plaque formation. Uninfected controls and wild type RSV controlplates were set up for comparison. After five to six days, each platewas overlayed with an additional 2 ml of 0.8% agarose in growth mediumsupplemented with 50 μg/ml neutral red. Following an overnightincubation in growth conditions, plaques were scored and picked for asecond round of purification.

[0385] Amplification of Clonal MARMs:

[0386] Following a second round of plaque purification three clones fromeach isolate were expanded for production of high titer viral stocks.Individual plaques were picked with the aid of a pipet tip and incubatedin 0.2 ml of fresh medium at 4° C. overnight to elute the virus. Eachaliquot was used to infect HEp-2 cells in flat bottom 24-well plates(2.5×10⁵ cells/well-seeded day before) in the presence of 4 μg/ml A4B4.After five days the entire well content was used to infect HEp-2 cellsin the presence of 4 μg/ml of A4b4 in 24-well plates. Each inoculum wassubsequently expanded in the presence of 4 μg/ml of A4b4 to a T-25 flask(6.5×10 cells/flask-seeded day before) and 5 mls of high titer virusstock were produced. MARM stocks were frozen as described above.

[0387] F Protein cDNA Synthesis and DNA Sequencing:

[0388] In order to determine the nucleotide sequence of an approximately800 nucleotide region of the F protein gene thought to contain theepitope for SYNAGIS®, First strand cDNA synthesis was performed withmRNAs isolated from MARM-infected HEp-2 cells at 4 days post-infection.RSV infected Hep2 cells were lysed in 150 μl of lysis buffer providedwith the mRNA capture kit. Biotinylated oligo dT was diluted 1:10 withnuclease free H₂O and 4 μl was added to each lysate. Samples wereincubated 10 minutes at 42° C. to allow the oligo dT to anneal to themRNA. A 50 μl aliquot of the lysate was transferred to a streptavidincoated PCR tube and incubated for three minutes at 37° C. The lysateswere removed from the PCR tubes and discarded. The RNA captured in thetubes was washed three times with 200 μl of wash buffer.

[0389] RT reactions were performed using reagents from the First StrandcDNA kit (Roche Molecular Biochemicals). A master mix was prepared sothat each reaction contained 5 μl 10×buffer, 5 μl dNTPs, 10 μl MgCl₂, 1μl gelatin, 2 μl RNase Inhibitor, 2 μl AMV-RT, in a final volume of 50μl. Fifty microliter aliquots of the master mix were transferred to thePCR tubes containing the captured mRNA. Samples were placed in athermalcycler and incubated for two hours at 42° C. The cDNA reactionmix was then removed from the PCR tubes and discarded. The cDNA capturedin the PCR tubes was washed with 200 μl of wash buffer To obtain enoughof the RSV F protein gene for sequence analysis, the cDNA was subjectedto PCR using gene specific primers. Each reactions contained 10 mMTris-HCl, pH 8.3, 50 mM KCl, 2.5 mM MgCl₂, 200 μM dNTPs, 125 ng of eachforward (5′ AGTGTCTTAACCAGCAAAGTGTTAGA 3′; SEQ ID NO: 258) and reverse(5′ TCATTGACTTGAGATATTGATGCATC 3′; SEQ ID NO: 259) primer, and 2.5 unitsof Taq polymerase (PE Biosystems) in a final volume of 50 μl. Thetemperature profile for all reactions was 95° C. for 2 min, followed by40 cycles of 95° C. for 30 sec., 55° C. for 45 sec, 72° C. for 45 sec,with a final extension at 72° C. for 10 minutes.

[0390] All PCR products were separated by electrophoresis on a 2%agarose gel in 1×TBE and visualized by ethidium bromide fluorescence.

[0391] PCR products were purified using Qiaquick spin columns (Qiagenand sequenced using the Big Dye-terminator PRISM kit (Applied Biosystems(ABI)). The reactions contained 70 ng of PCR product as template, 3pmols of primer, and 8.0 μl of the PRISM dye-terminator reaction mix ina final volume of 20 μl. The reactions were subjected to thermal cyclingaccording to ABI's dye-terminator sequencing instructions.Unincorporated dyes were removed from the extension products usingCentri-Sep spin columns (Princeton 30 Separations). Extension productswere dried in a Savant Speed Vac and then dissolved in 10 μl HiDiFormamide (ABI) loading buffer. Samples were applied by electrophoresisin an ABI 3100 automated sequencer. Sequence data collected by thesequencer was analyzed using Lasargene (DNA Star).

[0392] Immunofluorescence Assay (IFA):

[0393] Cells infected with RSV isolates were tested for anti-RSV bindingby A4B4, SYNAGIS® and Control Pan RSV MAb Pool, as follows. Four to fiveday RSV-infected HEp-2 cultures were mixed with PBS and centrifuged at300×g for 5 minutes at room temperature. Pellets were resuspended in asmall volume of PBS for analysis. Five to ten micro liters of each cellsuspension were spotted per 5 mm well on acetone washed 12-well HTCsupercured glass slides and allowed to air dry. Slides were fixed incold (−20° C.) acetone for 10 minutes. Reactions were blocked byspotting 10 μl of 1% BSA in PBS into each well and incubating for 10minutes at room temperature. Slides were washed three times in1×PBS/0.1% Tween-20 and air-dried. Ten micro liters of each primaryantibody reagent diluted to 250 ng/ml in blocking buffer were spottedper well and reactions were incubated in a humidified 37° C. environmentfor 30 minutes. Slides were then washed for 1 minute in three changes of1×PBS/0.1% Tween-20 and were air-dried. Ten micro liters of appropriatesecondary conjugated antibody reagent diluted to 250 ng/ml in blockingbuffer were added to each respective well and reactions were incubatedin a humidified 37° C. environment for an additional 30 minutes. Slideswere then washed for 1 minute in three changes of 1×PBS/0.1% Tween-20.Five micro liters of 50% glycerol in PBS, 10 mM Tris, pH 8.0, 1 mM EDTAwere spotted in each reaction well, and slides were mounted with coverslips. Each reaction well was subsequently analyzed by fluorescencemicroscopy at 200×power using a B-2A filter (EX 450-490 nm). Positivereactions were scored against an auto fluorescent background obtainedwith unstained cells or cells stained with secondary reagent alone.

[0394] RSV positive reactions were characterized by bright fluorescencepunctuated with small inclusions in the cytoplasm of infected cells.

[0395] Microneutralization Assay:

[0396] The procedure used here is described in Johnson et al., 1999, J.Infectious Diseases 180:35-40, the disclosure of which is herebyincorporated by reference in its entirety. Briefly, antibody dilutionswere made in triplicate using a 96-well plate. Ten TCID₅₀ of RSV A MARMSwere incubated with serial dilutions of the antibody to be tested for 2hours at 37° C. in the wells of a 96-well plate. The antibodies used inthe assay comprised the heavy and light chain of SYNAGIS®, the heavy andlight chain of A4b4, the combination of the heavy and light chain ofSYNAGIS® and A4b4, the SYNAGIS® heavy chain or light chain with a pointmutation in light chain or heavy chain, or A4b4 heavy chain with pointmutations in light chain. Mab 13/19 was used as a positive control inthe assay. RSV susceptible HEp-2 cells (2.5×10⁴) were then added to eachwell and cultured for 5 days at 37° C. in 5% CO₂. After 5 days, themedium was aspirated and cells were washed and fixed to the plates with80% methanol and 20% PBS. RSV replication was then determined by Fprotein expression. Fixed cells were incubated with a biotin-conjugatedanti-F protein monoclonal antibody (pan F protein, C-site-specific MAb133-1H) washed and horseradish peroxidase conjugated avidin was added tothe wells. The wells were washed again and turnover of substrate TMB(thionitrobenzoic acid) was measured at 450 nm. The results from theassay for the monoclonal antibodies are shown in Table 22 infra.

Results and Discussion

[0397] The binding activity of A4B4 was tested by IFA against a panel ofRSV A/Long MARMs obtained by three rounds of selection on HEp-2 cells inthe presence of SYNAGIS®. A pool of monoclonal antibodies against thefusion, glycoprotein, and nuclear proteins of RSV (Control Pan RSV MAbPool) was used as a positive control for detection of RSV. Subtyping ofRSV MARMs was performed with two monoclonal antibodies that distinguishtype A and type B glycoproteins. As summarized in Table 20, lack ofbinding activity by A4B4 and SYNAGIS® was demonstrated in MARMs.Contrasting with lack of binding by A4B4 and SYNAGIS®, binding of theControl Pan RSV MAb Pool was demonstrated for all MARMs tested. BothMARMs were classified as RSV type A. Wild type RSV A/Long infected HEp-2cells bound to A4B4 and SYNAGIS®, Pan RSV MAb Pool, and RSV Type A MAb,but failed to react with RSV Type B MAb, as expected.

[0398] DNA sequencing analysis of an approximately 800-nucleotide regionof the RSV F protein cDNA encompassing the proposed A4B4 epitoperevealed mutation at the amino-acid level at position 272 and 276. Table21 shows the amino acid change in isolates sequenced to date. Althoughthe entire nucleotide sequence of RSV MARMs F protein has not beendetermined, these results suggest that amino acid 272 and 276 is acritical residue in modulating the binding of A4B4 to its epitope.

[0399] The ability of various monoclonal antibodies to neutralize thereplication of a RSV A MARM was determined. As shown in Table 22, theability of the monoclonal antibodies to neutralize the replication of aRSV MARM varied depending upon the amino acid sequence of the heavychain (HC) and light chain (LC) of the antibody. TABLE 20Characterization of anti-RSV Binding Activity by A4B4, Synagis ®,Control Pan RSV MAb Pool (anti-F, G, N proteins), anti-RSV Type A MAb,and anti-RSV Type B MAb by Immunofluorescence Assay (IFA) on RSV A/LongMARMs. Reactivity w/ Reactivity w/ Reactivity w/ RSV MARM Reactivity w/Anti-RSV Anti-RSV Anti-RSV Reactivity sample Synagis ® MAb Pool type AMAb type B MAb A4B4 MARM #6 − + + − − MARM #9 − + + − − MARM #10 − + + −− MARM #11 − + + − − Wt + + + − +

[0400] TABLE 21 Amino acid sequence of a region of the wild type RSVA/Long and MARMs F Protein. Isolate Amino Acid Sequence (Positionnumber) Wild type 266 267 268 269 270 271 272 273 274 275 276 277 278279 RSV I T N D Q K K L M S N N V Q MARM I T N D Q K E L M S Y N V Q #10MARM I T N D Q K E L M S N N V Q #6 MARM I T N D Q K E L M S N N V Q #9MARM I T N D Q K E L M S N N V Q #11 MARM I T N D Q K E L M S N N V Q#14

[0401] TABLE 22 MARM analysis by Microneutralization Assay RSVMicroneutralization Assay On RSV A MARMs to A4B4 with RSV MonoclonalAntibody K272E, N276Y Antibody HC/LC MARM to MARM 10 NoteSYNAGIS ®/SYNAGIS ® − purified Ab A4B4/A4B4 − purified Ab A4b4/SYNAGIS ®− purified Ab SYNAGIS ®/A4B4 − purified Ab A4B4/L1FR − purified AbA4B4/L1FR S28R − purified Ab A4B4/L1FR S28R, S52F − purified AbA4B4/L1FR-28R, 52F, 93F − purified Ab A4B4/A4B428S − purified AbA4B4/A4B452S − purified Ab Mab 13/19 + purified Ab

14. EXAMPLE IN VITRO ISOLATION OF A4B4 SPECIFIC MONOCLONAL ANTIBODYRESISTANT MUTANTS (MARM) OF MARM B9(MARM OF RSV STRAIN A/LONG TOSYNAGIS)

[0402] This example demonstrates that MARMs can be isolated from MARM ofRSVA long to Synagis (MARM B9) upon multiple rounds of selection in thepresence of A4B4

Materials

[0403] A4B4 (Lot #524-9, 3.57 mg /ml), Control Pan RSV MAb Pool(Chemicon MAB858-4, a blend of three MAbs anti-F, G, and N proteins),anti-RSV Type A MAb (Chemicon MAB858-1), and anti-RSV Type B MAb(Chemicon MAB858-2) were used in this study. Secondary detectingreagents were either Alexa™488 conjugated Goat F (ab′)₂ anti-Mouse orHuman IgG (H+L). MARM B9 (1.78×10⁶ TCID₅₀/ml) was used here. HEp-2 cellswere propagated in EMEM supplemented with 10% FBS and 2 mM L-Gln in a37° C., 5% CO₂ environment. Messanger RNA was purified from the infectedcells using the mRNA Capture Kit (Roche). The mRNA samples were used tomake cDNA using the reagents from the cDNA First Strand Reacition Kit(Roche), followed by amplification of the RSV F protein gene by thePolymerase Chain Reaction (PCR) using gene specific primers.

Methods

[0404] Selection of MARMs:

[0405] 4×10⁵ HEp-2 cells were seeded per well of a 24-well plate ingrowth medium (EMEM, 10% FBS, 2 mM L-Gln) and incubated overnight ingrowth conditions (37° C., 5% CO₂). 44 wells were seeded for individualMARM selection. Prior to infection, a fresh vial of virus bank MARM B9was thawed rapidly at 37° C. and the virus titer adjusted to 6×10⁵pfu/ml in HEp-2 growth medium. A4B4 was added to the virus inoculum at afinal concentration of 2 μg/ml, and the mixture was incubated at 37° C.for one hour. An aliquot of RSV was incubated with an irrelevant HumanIgG1 Mab (MEDI-507) and was used as a negative control. An uninfectedcontrol well was also set up for each plate. Cells were washed once withfresh medium and were overlayed with 100 μl of RSV A/Long virusstock/MAb mixture. Cells were incubated for four hours in growthconditions followed by addition of 1 ml of growth medium to each well.Cytopathic effect (CPE) was monitored on a daily basis by lightmicroscopy. Following seven days of selection the contents of each wellwere supplemented with a further 4 μg/ml of A4B4 or irrelevant HumanIgG1 Mab, and were used to infect freshly seeded HEp-2 cells (4×10⁵cells/ well of 24-well plate). After a further seven days of selectionthe process was repeated one more time for a total of three rounds ofselection. The contents of wells showing clear CPE were mixed 1:1 with50% sucrose solution and immediately frozen at −80° C.

[0406] Plague Purification of MARMs:

[0407] After the third round of selection, the contents of independentwells were used for plaque purification of MARMs. A fresh vial of MARMstock (was frozen after third round of selection) was thawed at roomtemperature and was diluted 1:10, 1:100, and 1:1000 in growth medium andincubated with 4 μg/ml of A4B4 for one hour prior to infection. 0.5 mlof virus inoculum was overlayed on monolayers of HEp-2 cells in 6 wellplates (5×10⁵ cells/well) and incubated for 4 hours in growthconditions. Following infection the inoculum was carefully aspirated andcells were overlayed with 2 ml growth medium supplemented with 0.8% lowmelting temperature agarose (Gibco BRL), and containing 4 μg/ml of A4B4.Dishes were returned to the incubator after complete solidification ofagarose and monitored daily for plaque formation. Uninfected controlsand MARM B9 RSV control plates were set up for comparison. After five tosix days, each plate was overlayed with an additional 2 ml of 0.8%agarose in growth medium supplemented with 50 μg/ml neutral red.Following an overnight incubation in growth conditions, plaques werescored and picked for a second round of purification.

[0408] Amplification of Clonal MARMs:

[0409] Following a second round of plaque purification three clones fromisolate were expanded for production of high titer viral stocks.Individual plaques were picked with the aid of a pipet tip and incubatedin 0.2 ml of fresh medium at 4° C. overnight to elute the virus. Eachaliquot was used to infect HEp-2 cells in flat bottom 24-well plates(2.5×10⁵ cells/well- seeded day before) in the presence of 4 μg/ml A4B4.After five days the entire well content was used to infect HEp-2 cellsin the presence of 4 μg/ml of A4B4 in 24-well plates. Each inoculum wassubsequently expanded in the presence of 4 μg/ml of A4B4 to a T-25 flask(6.5×10⁵ cells/flask-seeded day before) and 5 mls of high titer virusstock were produced. MARM stocks were frozen as described above.

[0410] F Protein cDNA Synthesis and DNA Sequencing:

[0411] In order to determine the nucleotide sequence of an approximately800 nucleotide region of the F protein gene thought to contain theepitope for SYNAGIS®, First strand cDNA synthesis was performed withmRNAs isolated from MARM-infected HEp-2 cells at 4 days post-infection.RSV infected Hep2 cells were lysed in 150 μl of lysis buffer providedwith the mRNA capture kit. Biotinylated oligo dT was diluted 1:10 withnuclease free H₂O and 4 μl was added to each lysate. Samples wereincubated 10 minutes at 42° C. to allow the oligo dT to anneal to themRNA. A 50 μl aliquot of the lysate was transferred to a streptavidincoated PCR tube and incubated for three minutes at 37° C. The lysateswere removed from the PCR tubes and discarded. The RNA captured in thetubes was washed three times with 200 μl of wash buffer.

[0412] RT reactions were performed using reagents from the First StrandcDNA kit (Roche Molecular Biochemicals). A master mix was prepared sothat each reaction contained 5 μl 10×buffer, 5 μl dNTPs, 10 μl MgCl₂, 1μl gelatin, 2 μl RNase Inhibitor, 2 μl AMV-RT, in a final volume of 50μl. Fifty microliter aliquots of the master mix were transferred to thePCR tubes containing the captured mRNA. Samples were placed in athermalcycler and incubated for two hours at 42° C. The cDNA reactionmix was then removed from the PCR tubes and discarded. The cDNA capturedin the PCR tubes was washed with 200 μl of wash buffer To obtain enoughof the RSV F protein gene for sequence analysis, the cDNA was subjectedto PCR using gene specific primers. Each reactions contained 10 mMTris-HCl, pH 8.3, 50 mM KCl, 2.5 mM MgCl₂, 200 FM dNTPs, 125 ng of eachforward (5′ AGTGTCTTAACCAGCAAAGTGTTAGA 3′; SEQ ID NO: 258) and reverse(5′ TCATTGACTTGAGATATTGATGCATC 3′; SEQ ID NO: 259) primer, and 2.5 unitsof Taq polymerase (PE Biosystems) in a final volume of 50 μl. Thetemperature profile for all reactions was 95° C. for 2 min, followed by40 cycles of 95° C. for 30 sec., 55° C. for 45 sec, 72° C. for 45 sec,with a final extension at 72° C. for 10 minutes.

[0413] All PCR products were separated by electrophoresis on a 2%agarose gel in 1×TBE and visualized by ethidium bromide fluorescence.

[0414] PCR products were purified using Qiaquick spin columns (Qiagenand sequenced using the Big Dye-terminator PRISM kit (Applied Biosystems(ABI)). The reactions contained 70 ng of PCR product as template, 3pmols of primer, and 8.0 μl of the PRISM dye-terminator reaction mix ina final volume of 20 μl. The reactions were subjected to thermal cyclingaccording to ABI's dye-terminator sequencing instructions.Unincorporated dyes were removed from the extension products usingCentri-Sep spin columns (Princeton Separations). Extension products weredried in a Savant Speed Vac and then dissolved in 10 μl HiDi Fornamide(ABI) loading buffer. Samples were applied by electrophoresis in an ABI3100 automated sequencer. Sequence data collected by the sequencer wasanalyzed using Lasargene (DNA Star).

[0415] Immunofluorescence Assay (IFA:

[0416] Cells infected with RSV isolates were tested for anti-RSV bindingby A4B4, SYNAGIS® and Control Pan RSV MAb Pool, as follows. Four to fiveday RSV-infected HEp-2 cultures were mixed with PBS and centrifuged at300×g for 5 minutes at room temperature. Pellets were resuspended in asmall volume of PBS for analysis. Five to ten micro liters of each cellsuspension were spotted per 5 mm well on acetone washed 12-well HTCsuper cured glass slides and allowed to air dry. Slides were fixed incold (−20° C.) acetone for 10 minutes. Reactions were blocked byspotting 10 μl of 1% BSA in PBS into each well and incubating for 10minutes at room temperature. Slides were washed three times in 1×PBS/0.1% Tween-20 and air-dried. Ten micro liters of each primary antibodyreagent diluted to 250 ng/ml in blocking buffer were spotted per welland reactions were incubated in a humidified 37° C. environment for 30minutes. Slides were then washed for 1 minute in three changes of1×PBS/0.1% Tween-20 and were air-dried. Ten micro liters of appropriatesecondary conjugated antibody reagent diluted to 250 ng/ml in blockingbuffer were added to each respective well and reactions were incubatedin a humidified 37° C. environment for an additional 30 minutes. Slideswere then washed for 1 minute in three changes of 1×PBS/0.1% Tween-20.Five micro liters of 50% glycerol in PBS, 10 mM Tris, pH 8.0, 1 mM EDTAwere spotted in each reaction well, and slides were mounted with coverslips. Each reaction well was subsequently analyzed by fluorescencemicroscopy at 200×power using a B-2A filter (EX 450-490 nm). Positivereactions were scored against an auto fluorescent background obtainedwith unstained cells or cells stained with secondary reagent alone.

[0417] RSV positive reactions were characterized by bright fluorescencepunctuated with small inclusions in the cytoplasm of infected cells.

[0418] Microneutralization Assay:

[0419] The procedure used here is described in Johnson et al., 1999, J.Infectious Diseases 180:35-40, the disclosure of which is herebyincorporated by reference in its entirety. Briefly, antibody dilutionswere made in triplicate using a 96-well plate. Ten TCID₅₀ of RSV A MARMSwere incubated with serial dilutions of the antibody to be tested for 2hours at 37° C. in the wells of a 96-well plate. The antibodies used inthe assay comprised the heavy chain of SYNAGIS® or the heavy chain ofA4B4, and the light chain of SYNAGIS®, the light chain of A4B4, thelight chain of A4B4 with a point mutation, the light chain of L1FR orthe light chain of L1FR with point mutations. Mab 13/19 was used as apositive control in the assays. RSV susceptible HEp-2 cells (2.5×10⁴)were then added to each well and cultured for 5 days at 37° C. in 5%CO₂. After 5 days, the medium was aspirated and cells were washed andfixed to the plates with 80% methanol and 20% PBS. RSV replication wasthen determined by F protein expression. Fixed cells were incubated witha biotin-conjugated anti-F protein monoclonal antibody (pan F protein,C-site-specific MAb 133-1H) washed and horseradish peroxidase conjugatedavidin was added to the wells. The wells were washed again and turnoverof substrate TMB (thionitrobenzoic acid) was measured at 450 nm. Theresults from the assay for the monoclonal antibodies are shown in Table25 infra.

Results and Discussion

[0420] The binding activity of A4B4 was tested by IFA against a MARMobtained by three rounds of selection on HEp-2 cells in the presence ofA4B4. A pool of monoclonal antibodies against the fusion, glycoprotein,and nuclear proteins of RSV (Control Pan RSV MAb Pool) was used as apositive control for detection of RSV. Subtyping of RSV MARMs wasperformed with two monoclonal antibodies that distinguish type A andtype B glycoproteins. As summarized in Table 23, lack of bindingactivity by A4B4 and SYNAGIS® was demonstrated in both MARMs.Contrasting with lack of binding by A4B4 and SYNAGIS®, binding of theControl Pan RSV MAb Pool was demonstrated for all MARMs tested. BothMARMs were classified as RSV type A. Wild type RSV A/Long infected HEp-2cells bound to A4B4 and SYNAGIS®, Pan RSV MAb Pool, and RSV Type A MAb,but failed to react with RSV Type B MAb, as expected.

[0421] DNA sequencing analysis of an approximately 800 nucleotide regionof the RSV F protein cDNA encompassing the proposed A4B4 epitoperevealed mutation at the amino-acid level at position 272 and 262. Table24 shows the amino acid change in isolates sequenced to date. Althoughthe entire nucleotide sequence of RSV MARMs F protein has not beendetermined, these results suggest that amino acid 272 and 262 is acritical residue in modulating the binding of A4B4 to its epitope.

[0422] The ability of various monoclonal antibodies to neutralize thereplication of RSV A MARMs was determined. As shown in Table 25, theability of the monoclonal antibodies to neutralize the replication ofRSV MARMs varied depending upon the amino acid sequence of the heavychain (HC) and light chain (LC) of the antibody. TABLE 23Characterization of anti-RSV Binding Activity by A4B4, SYNAGIS ®,Control Pan RSV MAb Pool (anti-F, G, N proteins), anti-RSV Type A MAb,and anti-RSV Type B MAb by Immunofluorescence Assay (IFA) on RSV A/LongMARMs. Reactivity w/ Reactivity w/ Reactivity w/ RSV MARM Reactivity w/Anti-RSV Anti-RSV Anti-RSV Reactivity sample Synagis ® MAb Pool type AMAb type B MAb A4B4 MARM #13 − + + − − B9 − + + − + Wt + + + − −

[0423] TABLE 24 Amino acid sequence of a region of the wild type RSVA/Long and MARMs F Protein. Isolate Amino Acid Sequence (Positionnumber) Wild type 262 266 267 268 269 270 271 272 273 274 275 276 277278 279 RSV Long A N I T N D Q K K L M S N N V Q MARM 262 266 267 268269 270 271 272 273 274 275 276 277 278 279 #13 K I T N D Q K Q L M S NN V Q

[0424] TABLE 25 MARM analysis by Microneutralization Assay RSVMicroneutralization Assay On RSV A MARMs to A4B4 with RSV MonoclonalAntibody. K272Q, N262K MARM to SYNAGIS ® & Antibody HC/LC A4B4 MARM 13Note SYNAGIS ®/SYNAGIS ® − purified Ab A4B4/A4B4 − purified AbA4B4/SYNAGIS ® − purified Ab SYNAGIS ®/A4B4 − purified Ab A4B4/L1FR −purified Ab A4B4/L1FR S28R − purified Ab A4B4/L1FR S28R, S52F − purifiedAb A4B4/L1FR-28R, 52F, 93F − purified Ab A4B4/A4B428S − purified AbA4B4/A4B452S − purified Ab Mab 13/19 + purified Ab

15. EXAMPLE CLINICAL TRIALS

[0425] Antibodies of the invention or fragments thereof tested in invitro assays and animal models may be further evaluated for safety,tolerance and pharmacokinetics in groups of normal healthy adultvolunteers. The volunteers are administered intramuscularly,intravenously or by a pulmonary delivery system a single dose of 0.5mg/kg, 3 mg/kg, 5 mg/kg, 10 mg/kg or 15 mg/kg of an antibody or fragmentthereof which immunospecifically binds to a RSV antigen. Each volunteeris monitored at least 24 hours prior to receiving the single dose of theantibody or fragment thereof and each volunteer will be monitored for atleast 48 hours after receiving the dose at a clinical site. Thenvolunteers are monitored as outpatients on days 3, 7, 14, 21, 28, 35,42, 49, and 56 postdose.

[0426] Blood samples are collected via an indwelling catheter or directvenipuncture using 10 ml red-top Vacutainer tubes at the followingintervals: (1) prior to administering the dose of the antibody orantibody fragment; (2) during the administration of the dose of theantibody or antibody fragment; (3) 5 minutes, 10 minutes, 15 minutes, 20minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 24hours, and 48 hours after administering the dose of the antibody orantibody fragment; and (4) 3 days, 7 days 14 days, 21 days, 28 days, 35days, 42 days, 49 days, and 56 days after administering the does of theantibody or antibody fragment. Samples are allowed to clot at roomtemperature and serum will be collected after centrifugation.

[0427] The antibody or antibody fragment is partially purified from theserum samples and the amount of antibody or antibody fragment in thesamples will be quantitated by ELISA. Briefly, the ELISA consists ofcoating microtiter plates overnight at 4° C. with an antibody thatrecognizes the antibody or antibody fragment administered to thevolunteer. The plates are then blocked for approximately 30 minutes atroom temperature with PBS-Tween-0.5% BSA. Standard curves areconstructed using purified antibody or antibody fragment, notadministered to a volunteer. Samples are diluted in PBS-Tween-BSA. Thesamples and standards are incubated for approximately 1 hour at roomtemperature. Next, the bound antibody is treated with a labeled antibody(e.g., horseradish peroxidase conjugated goat-anti-human IgG) forapproximately 1 hour at room temperature. Binding of the labeledantibody is detected, e.g., by a spectrophotometer.

[0428] The concentration of antibody or antibody fragment levels in theserum of volunteers are corrected by subtracting the predose serum level(background level) from the serum levels at each collection intervalafter administration of the dose. For each volunteer the pharmacokineticparameters are computed according to the model-independent approach(Gibaldi et al., eds., 1982, Pharmacokinetics, 2^(nd) edition, MarcelDekker, New York) from the corrected serum antibody or antibody fragmentconcentrations.

[0429] Equivalents

[0430] Those skilled in the art will recognize, or be able to ascertainusing no more than routine experimentation, many equivalents to thespecific embodiments of the invention described herein. Such equivalentsare intended to be encompassed by the following claims.

[0431] All publications, patents and patent applications mentioned inthis specification are herein incorporated by reference into thespecification to the same extent as if each individual publication,patent or patent application was specifically and individually indicatedto be incorporated herein by reference.

1 259 1 7 PRT Homo sapiens 1 Thr Ser Gly Met Ser Val Gly 1 5 2 16 PRTHomo sapiens 2 Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser LeuLys Ser 1 5 10 15 3 10 PRT Homo sapiens 3 Ser Met Ile Thr Asn Trp TyrPhe Asp Val 1 5 10 4 10 PRT Homo sapiens 4 Lys Cys Gln Leu Ser Val GlyTyr Met His 1 5 10 5 7 PRT Homo sapiens 5 Asp Thr Ser Lys Leu Ala Ser 15 6 9 PRT Homo sapiens 6 Phe Gln Gly Ser Gly Tyr Pro Phe Thr 1 5 7 120PRT Homo sapiens 7 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val LysPro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe SerLeu Ser Thr Ser 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro GlyLys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys AspTyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr SerLys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala AspThr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile Thr Asn Trp Tyr PheAsp Val Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 1208 106 PRT Homo sapiens misc_feature VL Domain 8 Asp Ile Gln Met Thr GlnSer Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr IleThr Cys Lys Cys Gln Leu Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Lys LeuAla Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly GlyThr Lys Leu Glu Ile Lys 100 105 9 120 PRT Homo sapiens misc_feature VHDomain 9 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser ThrAla 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala LeuGlu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn ProSer 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn GlnVal 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala ThrTyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile Thr Asn Phe Tyr Phe Asp Val TrpGly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 10 7 PRTHomo sapiens 10 Thr Ala Gly Met Ser Val Gly 1 5 11 106 PRT Homo sapiens11 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 510 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val Gly Tyr Met 2025 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 3540 45 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 5055 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 6570 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 12 10 PRT Homosapiens 12 Ser Met Ile Thr Asn Phe Tyr Phe Asp Val 1 5 10 13 106 PRTHomo sapiens 13 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala SerVal Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser ValGly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys LeuLeu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Ala Ser Gly Val Pro Ser Arg PheSer Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser LeuGln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Phe Ser GlyTyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 10514 10 PRT Homo sapiens 14 Ser Ala Ser Ser Ser Val Gly Tyr Met His 1 5 1015 7 PRT Homo sapiens 15 Asp Thr Phe Lys Leu Ala Ser 1 5 16 9 PRT Homosapiens 16 Phe Gln Phe Ser Gly Tyr Pro Phe Thr 1 5 17 120 PRT Homosapiens 17 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro ThrGln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu SerThr Pro 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys AlaLeu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys His Tyr AsnPro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys AsnGln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr AlaThr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp ValTrp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 18 7 PRTHomo sapiens 18 Thr Pro Gly Met Ser Val Gly 1 5 19 16 PRT Homo sapiens19 Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser Leu Lys Asp 1 510 15 20 10 PRT Homo sapiens 20 Asp Met Ile Phe Asn Phe Tyr Phe Asp Val1 5 10 21 106 PRT Homo sapiens misc_feature VL Domain 21 Asp Ile Gln MetThr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg ValThr Ile Thr Cys Ser Leu Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp TyrGln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr PheTyr Leu Ser Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser GlyThr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp PheAla Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe GlyGly Gly Thr Lys Val Glu Ile Lys 100 105 22 10 PRT Homo sapiens 22 SerLeu Ser Ser Arg Val Gly Tyr Met His 1 5 10 23 7 PRT Homo sapiens 23 AspThr Phe Tyr Leu Ser Ser 1 5 24 120 PRT Homo sapiens misc_feature VHDomain 24 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro ThrGln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu SerThr Pro 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys AlaLeu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Gly Lys Lys His Tyr AsnPro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys AsnGln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr AlaThr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp ValTrp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 25 16PRT Homo sapiens 25 Asp Ile Trp Trp Asp Gly Lys Lys His Tyr Asn Pro SerLeu Lys Asp 1 5 10 15 26 106 PRT Homo sapiens 26 Asp Ile Gln Met Thr GlnSer Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr IleThr Cys Ser Leu Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Arg Gly LeuPro Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly GlyThr Lys Val Glu Ile Lys 100 105 27 7 PRT Homo sapiens 27 Asp Thr Arg GlyLeu Pro Ser 1 5 28 120 PRT Homo sapiens 28 Gln Val Thr Leu Arg Glu SerGly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr CysThr Phe Ser Gly Phe Ser Leu Ser Thr Pro 20 25 30 Gly Met Ser Val Gly TrpIle Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile TrpTrp Asp Gly Lys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu ThrIle Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val ThrAsn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp MetIle Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr ValThr Val Ser Ser 115 120 29 10 PRT Homo sapiens 29 Asp Met Ile Phe AsnTrp Tyr Phe Asp Val 1 5 10 30 106 PRT Homo sapiens 30 Asp Ile Gln MetThr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg ValThr Ile Thr Cys Ser Pro Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp TyrGln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr MetArg Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser GlyThr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp PheAla Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe GlyGly Gly Thr Lys Val Glu Ile Lys 100 105 31 10 PRT Homo sapiens 31 SerPro Ser Ser Arg Val Gly Tyr Met His 1 5 10 32 7 PRT Homo sapiens 32 AspThr Met Arg Leu Ala Ser 1 5 33 120 PRT Homo sapiens 33 Gln Val Thr LeuArg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu ThrLeu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Pro 20 25 30 Gly Met SerVal Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu AlaAsp Ile Trp Trp Asp Gly Lys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys AspArg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val LeuLys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys AlaArg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln 100 105 110 GlyThr Thr Val Thr Val Ser Ser 115 120 34 106 PRT Homo sapiens 34 Asp IleGln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 AspArg Val Thr Ile Thr Cys Ser Leu Ser Ser Arg Val Gly Tyr Met 20 25 30 HisTrp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 AspThr Phe Lys Leu Ser Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 GlySer Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 35 7 PRT Homo sapiens 35Asp Thr Phe Lys Leu Ser Ser 1 5 36 120 PRT Homo sapiens 36 Gln Val ThrLeu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr LeuThr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly MetSer Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp LeuAla Asp Ile Trp Trp Asp Gly Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu LysAsp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 ValLeu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 CysAla Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser 115 120 37 16 PRT Homo sapiens 37 AspIle Trp Trp Asp Gly Lys Lys Asp Tyr Asn Pro Ser Leu Lys Asp 1 5 10 15 38106 PRT Homo sapiens 38 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser SerArg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Ser Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys100 105 39 10 PRT Homo sapiens 39 Ser Ala Ser Ser Arg Val Gly Tyr MetHis 1 5 10 40 120 PRT Homo sapiens 40 Gln Val Thr Leu Arg Glu Ser GlyPro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys ThrPhe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp IleArg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp TrpAsp Gly Lys Lys Ser Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr IleSer Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr AsnMet Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met IlePhe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val ThrVal Ser Ser 115 120 41 16 PRT Homo sapiens 41 Asp Ile Trp Trp Asp GlyLys Lys Ser Tyr Asn Pro Ser Leu Lys Asp 1 5 10 15 42 106 PRT Homosapiens 42 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Leu Ser Ser Arg Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Met Tyr Gln Ser Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 437 PRT Homo sapiens 43 Asp Thr Met Tyr Gln Ser Ser 1 5 44 120 PRT Homosapiens 44 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro ThrGln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu SerThr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys AlaLeu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Gly Lys Lys Ser Tyr AsnPro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys AsnGln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr AlaThr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp ValTrp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 45 16PRT Homo sapiens 45 Asp Ile Trp Trp Asp Asp Lys Lys Ser Tyr Asn Pro SerLeu Lys Asp 1 5 10 15 46 106 PRT Homo sapiens 46 Asp Ile Gln Met Thr GlnSer Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr IleThr Cys Leu Pro Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Met Tyr GlnAla Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Phe Gln Phe Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly GlyThr Lys Leu Glu Ile Lys 100 105 47 10 PRT Homo sapiens 47 Leu Pro SerSer Arg Val Gly Tyr Met His 1 5 10 48 120 PRT Homo sapiens 48 Gln ValThr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 ThrLeu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 GlyMet Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 TrpLeu Ala Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser 50 55 60 LeuLys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105110 Gly Thr Thr Val Thr Val Ser Ser 115 120 49 106 PRT Homo sapiens 49Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 1015 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val Gly Tyr Met 20 2530 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Asp Thr Phe Phe Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 8590 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 50 7 PRT Homosapiens 50 Asp Thr Phe Phe Leu Asp Ser 1 5 51 120 PRT Homo sapiens 51Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 1015 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 2530 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 4045 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Ser Tyr Asn Pro Ser 50 5560 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 7075 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 8590 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 52 106 PRT Homosapiens 52 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Arg Tyr Gln Ser Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 537 PRT Homo sapiens 53 Asp Thr Arg Tyr Gln Ser Ser 1 5 54 106 PRT Homosapiens misc_feature VL Domain 54 Asp Ile Gln Met Thr Gln Ser Pro SerThr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys SerAla Ser Ser Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro GlyLys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Lys Leu Ala Ser GlyVal Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr LeuThr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr CysPhe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys LeuGlu Ile Lys 100 105 55 120 PRT Homo sapiens 55 Gln Val Thr Leu Arg GluSer Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu ThrCys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val GlyTrp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp IleTrp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg LeuThr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys ValThr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg AspMet Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Ala 100 105 110 Gly Thr ThrVal Thr Val Ser Ser 115 120 56 106 PRT Homo sapiens misc_feature VLDomain 56 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Phe Lys Leu Ala Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 57106 PRT Homo sapiens misc_feature VL Domain 57 Asp Ile Gln Met Thr GlnSer Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr IleThr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Tyr Lys GlnThr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr GluPhe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala ThrTyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly GlyThr Lys Leu Glu Ile Lys 100 105 58 7 PRT Homo sapiens 58 Asp Thr Tyr LysGln Thr Ser 1 5 59 106 PRT Homo sapiens misc_feature VL Domain 59 AspIle Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Arg Tyr Leu Ser Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 9095 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 60 106 PRT Homosapiens 60 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Phe Lys Leu Ala Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Phe TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 619 PRT Homo sapiens 61 Phe Gln Gly Ser Phe Tyr Pro Phe Thr 1 5 62 106 PRTHomo sapiens misc_feature VL Domain 62 Asp Ile Gln Met Thr Gln Ser ProSer Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr CysSer Ala Ser Ser Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys ProGly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Thr SerGly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe ThrLeu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr TyrCys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr LysLeu Glu Ile Lys 100 105 63 7 PRT Homo sapiens 63 Asp Thr Phe Lys Leu ThrSer 1 5 64 106 PRT Homo sapiens misc_feature VL Domain 64 Asp Ile GlnMet Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp ArgVal Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 25 30 His TrpTyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp ThrPhe Arg Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly SerGly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 AspPhe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 PheGly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 65 106 PRT Homo sapiens 65Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 1015 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 2530 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Asp Thr Phe Arg Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 8590 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 66 7 PRT Homosapiens 66 Asp Thr Phe Arg Leu Ala Ser 1 5 67 120 PRT Homo sapiens 67Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 1015 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 2530 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 4045 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser 50 5560 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 7075 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 8590 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Ala100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 68 106 PRT Homosapiens 68 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Tyr Arg His Ala Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 697 PRT Homo sapiens 69 Asp Thr Tyr Arg His Ser Ser 1 5 70 106 PRT Homosapiens 70 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Tyr Lys Gln Thr Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 71106 PRT Homo sapiens 71 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Leu Ser SerSer Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Phe His Arg Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys100 105 72 10 PRT Homo sapiens 72 Ser Leu Ser Ser Ser Val Gly Tyr MetHis 1 5 10 73 7 PRT Homo sapiens 73 Asp Thr Phe Phe His Arg Ser 1 5 74106 PRT Homo sapiens 74 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser SerArg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Leu Leu Leu Asp Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys100 105 75 7 PRT Homo sapiens 75 Asp Thr Leu Leu Leu Asp Ser 1 5 76 106PRT Homo sapiens 76 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser AlaSer Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser ArgVal Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro LysLeu Leu Ile Tyr 35 40 45 Asp Thr Ser Phe Leu Asp Ser Gly Val Pro Ser ArgPhe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser SerLeu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly SerGly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100105 77 7 PRT Homo sapiens 77 Asp Thr Ser Phe Leu Asp Ser 1 5 78 120 PRTHomo sapiens 78 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys ProThr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser LeuSer Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly LysAla Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp TyrAsn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser LysAsn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp ThrAla Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Thr Asn Phe Tyr Phe AspVal Trp Gly Ala 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 7910 PRT Homo sapiens 79 Asp Met Ile Thr Asn Phe Tyr Phe Asp Val 1 5 10 8010 PRT Homo sapiens 80 Lys Cys Gln Ser Ser Val Gly Tyr Met His 1 5 10 817 PRT Homo sapiens 81 Asp Thr Ser Tyr Leu Ala Ser 1 5 82 16 PRT Homosapiens 82 Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser Leu LysSer 1 5 10 15 83 10 PRT Homo sapiens 83 Asp Met Ile Thr Asn Trp Tyr PheAsp Val 1 5 10 84 10 PRT Homo sapiens 84 Lys Cys Gln Ser Arg Val Gly TyrMet His 1 5 10 85 7 PRT Homo sapiens 85 Asp Thr Ser Tyr Leu Ser Ser 1 586 16 PRT Homo sapiens 86 Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr AsnPro Ser Leu Lys Asp 1 5 10 15 87 10 PRT Homo sapiens 87 Lys Cys Gln LeuArg Val Gly Tyr Met His 1 5 10 88 7 PRT Homo sapiens 88 Asp Thr Lys LysLeu Ser Ser 1 5 89 10 PRT Homo sapiens 89 Lys Leu Gln Leu Ser Val GlyTyr Met His 1 5 10 90 7 PRT Homo sapiens 90 Asp Thr Phe Tyr Leu Ser Ser1 5 91 16 PRT Homo sapiens 91 Asp Ile Trp Trp Asp Asp Lys Lys His TyrAsn Pro Ser Leu Lys Ser 1 5 10 15 92 10 PRT Homo sapiens 92 Lys Leu GlnSer Ser Val Gly Tyr Met His 1 5 10 93 16 PRT Homo sapiens 93 Asp Ile TrpTrp Asp Asp Lys Lys His Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 94 10 PRTHomo sapiens 94 Ser Met Ile Phe Asn Trp Tyr Phe Asp Val 1 5 10 95 10 PRTHomo sapiens 95 Lys Leu Gln Ser Arg Val Gly Tyr Met His 1 5 10 96 7 PRTHomo sapiens 96 Asp Thr Phe Lys Leu Ser Ser 1 5 97 10 PRT Homo sapiens97 Ser Met Ile Phe Asn Phe Tyr Phe Asp Val 1 5 10 98 10 PRT Homo sapiens98 Lys Leu Gln Leu Arg Val Gly Tyr Met His 1 5 10 99 7 PRT Homo sapiens99 Asp Thr Phe Tyr Leu Ala Ser 1 5 100 16 PRT Homo sapiens 100 Asp IleTrp Trp Asp Gly Lys Lys Asp Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 101 10PRT Homo sapiens 101 Lys Leu Ser Leu Ser Val Gly Tyr Met His 1 5 10 1027 PRT Homo sapiens 102 Asp Thr Ser Lys Leu Pro Ser 1 5 103 16 PRT Homosapiens 103 Asp Ile Trp Trp Asp Gly Lys Lys Asp Tyr Asn Pro Ser Leu LysAsp 1 5 10 15 104 10 PRT Homo sapiens 104 Lys Leu Ser Ser Ser Val GlyTyr Met His 1 5 10 105 7 PRT Homo sapiens 105 Asp Thr Ser Gly Leu AlaSer 1 5 106 16 PRT Homo sapiens 106 Asp Ile Trp Trp Asp Gly Lys Lys HisTyr Asn Pro Ser Leu Lys Ser 1 5 10 15 107 10 PRT Homo sapiens 107 LysLeu Ser Ser Arg Val Gly Tyr Met His 1 5 10 108 7 PRT Homo sapiens 108Asp Thr Ser Gly Leu Pro Ser 1 5 109 16 PRT Homo sapiens 109 Asp Ile TrpTrp Asp Asp Lys Lys Ser Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 110 10 PRTHomo sapiens 110 Lys Leu Ser Leu Arg Val Gly Tyr Met His 1 5 10 111 16PRT Homo sapiens 111 Asp Ile Trp Trp Asp Asp Lys Lys Ser Tyr Asn Pro SerLeu Lys Ser 1 5 10 15 112 10 PRT Homo sapiens 112 Lys Cys Ser Leu SerVal Gly Tyr Met His 1 5 10 113 7 PRT Homo sapiens 113 Asp Thr Arg LysLeu Ala Ser 1 5 114 16 PRT Homo sapiens 114 Asp Ile Trp Trp Asp Gly LysLys Ser Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 115 10 PRT Homo sapiens115 Lys Cys Ser Ser Ser Val Gly Tyr Met His 1 5 10 116 7 PRT Homosapiens 116 Asp Thr Arg Gly Leu Ala Ser 1 5 117 10 PRT Homo sapiens 117Lys Cys Ser Leu Arg Val Gly Tyr Met His 1 5 10 118 7 PRT Homo sapiens118 Asp Thr Arg Lys Leu Pro Ser 1 5 119 10 PRT Homo sapiens 119 Lys CysSer Leu Arg Val Gly Tyr Met His 1 5 10 120 10 PRT Homo sapiens 120 SerLeu Ser Leu Ser Val Gly Tyr Met His 1 5 10 121 7 PRT Homo sapiens 121Asp Thr Met Lys Leu Ala Ser 1 5 122 10 PRT Homo sapiens 122 Ser Leu SerSer Ser Val Gly Tyr Met His 1 5 10 123 7 PRT Homo sapiens 123 Asp ThrSer Arg Leu Ala Ser 1 5 124 7 PRT Homo sapiens 124 Asp Thr Ser Leu LeuAla Ser 1 5 125 10 PRT Homo sapiens 125 Ser Leu Ser Leu Arg Val Gly TyrMet His 1 5 10 126 7 PRT Homo sapiens 126 Asp Thr Ser Leu Leu Asp Ser 15 127 10 PRT Homo sapiens 127 Ser Cys Gln Leu Ser Val Gly Tyr Met His 15 10 128 7 PRT Homo sapiens 128 Asp Thr Ser Lys Leu Asp Ser 1 5 129 10PRT Homo sapiens 129 Ser Cys Gln Ser Ser Val Gly Tyr Met His 1 5 10 13010 PRT Homo sapiens 130 Ser Cys Gln Ser Arg Val Gly Tyr Met His 1 5 10131 7 PRT Homo sapiens 131 Asp Thr Leu Lys Leu Asp Ser 1 5 132 10 PRTHomo sapiens 132 Ser Cys Gln Leu Arg Val Gly Tyr Met His 1 5 10 133 7PRT Homo sapiens 133 Asp Thr Leu Leu Leu Ala Ser 1 5 134 10 PRT Homosapiens 134 Ser Leu Gln Leu Ser Val Gly Tyr Met His 1 5 10 135 7 PRTHomo sapiens 135 Asp Thr Leu Lys Leu Ala Ser 1 5 136 10 PRT Homo sapiens136 Ser Leu Gln Ser Ser Val Gly Tyr Met His 1 5 10 137 7 PRT Homosapiens 137 Asp Thr Ser Lys Leu Ser Ser 1 5 138 10 PRT Homo sapiens 138Ser Leu Gln Ser Arg Val Gly Tyr Met His 1 5 10 139 7 PRT Homo sapiens139 Asp Thr Ser Lys Gln Ala Ser 1 5 140 10 PRT Homo sapiens 140 Ser LeuGln Leu Arg Val Gly Tyr Met His 1 5 10 141 7 PRT Homo sapiens 141 AspThr Ser Lys Gln Ser Ser 1 5 142 10 PRT Homo sapiens 142 Ser Cys Ser LeuSer Val Gly Tyr Met His 1 5 10 143 7 PRT Homo sapiens 143 Asp Thr SerTyr Leu Ala Ser 1 5 144 10 PRT Homo sapiens 144 Ser Cys Ser Ser Ser ValGly Tyr Met His 1 5 10 145 7 PRT Homo sapiens 145 Asp Thr Ser Tyr LeuSer Ser 1 5 146 10 PRT Homo sapiens 146 Ser Cys Ser Ser Arg Val Gly TyrMet His 1 5 10 147 7 PRT Homo sapiens 147 Asp Thr Ser Tyr Gln Ala Ser 15 148 10 PRT Homo sapiens 148 Ser Cys Ser Leu Arg Val Gly Tyr Met His 15 10 149 7 PRT Homo sapiens 149 Asp Thr Ser Tyr Gln Ser Ser 1 5 150 10PRT Homo sapiens 150 Lys Pro Ser Ser Arg Val Gly Tyr Met His 1 5 10 1517 PRT Homo sapiens 151 Asp Thr Met Tyr Gln Ala Ser 1 5 152 10 PRT Homosapiens 152 Lys Pro Ser Leu Arg Val Gly Tyr Met His 1 5 10 153 10 PRTHomo sapiens 153 Lys Pro Ser Ser Ser Val Gly Tyr Met His 1 5 10 154 7PRT Homo sapiens 154 Asp Thr Met Lys Gln Ala Ser 1 5 155 10 PRT Homosapiens 155 Lys Pro Ser Leu Ser Val Gly Tyr Met His 1 5 10 156 7 PRTHomo sapiens 156 Asp Thr Met Lys Gln Ser Ser 1 5 157 10 PRT Homo sapiens157 Lys Pro Gln Ser Arg Val Gly Tyr Met His 1 5 10 158 7 PRT Homosapiens 158 Asp Thr Met Tyr Leu Ala Ser 1 5 159 10 PRT Homo sapiens 159Lys Pro Gln Leu Arg Val Gly Tyr Met His 1 5 10 160 7 PRT Homo sapiens160 Asp Thr Met Tyr Leu Ser Ser 1 5 161 10 PRT Homo sapiens 161 Lys ProGln Ser Ser Val Gly Tyr Met His 1 5 10 162 7 PRT Homo sapiens 162 AspThr Met Lys Leu Ala Ser 1 5 163 10 PRT Homo sapiens 163 Lys Pro Gln LeuSer Val Gly Tyr Met His 1 5 10 164 7 PRT Homo sapiens 164 Asp Thr MetLys Leu Ser Ser 1 5 165 7 PRT Homo sapiens 165 Asp Thr Ser Lys Leu SerSer 1 5 166 10 PRT Homo sapiens 166 Ser Pro Ser Leu Arg Val Gly Tyr MetHis 1 5 10 167 7 PRT Homo sapiens 167 Asp Thr Ser Lys Leu Ser Ser 1 5168 10 PRT Homo sapiens 168 Ser Pro Ser Ser Ser Val Gly Tyr Met His 1 510 169 10 PRT Homo sapiens 169 Ser Pro Ser Leu Ser Val Gly Tyr Met His 15 10 170 7 PRT Homo sapiens 170 Asp Thr Arg Tyr Gln Ala Ser 1 5 171 10PRT Homo sapiens 171 Ser Pro Gln Ser Arg Val Gly Tyr Met His 1 5 10 1727 PRT Homo sapiens 172 Asp Thr Arg Lys Gln Ser Ser 1 5 173 10 PRT Homosapiens 173 Ser Pro Gln Leu Arg Val Gly Tyr Met His 1 5 10 174 7 PRTHomo sapiens 174 Asp Thr Arg Lys Leu Ala Ser 1 5 175 7 PRT Homo sapiens175 Asp Thr Arg Lys Leu Ser Ser 1 5 176 10 PRT Homo sapiens 176 Ser ProGln Ser Ser Val Gly Tyr Met His 1 5 10 177 10 PRT Homo sapiens 177 SerPro Gln Leu Ser Val Gly Tyr Met His 1 5 10 178 7 PRT Homo sapiens 178Asp Thr Arg Tyr Leu Ala Ser 1 5 179 10 PRT Homo sapiens 179 Lys Ala GlnSer Arg Val Gly Tyr Met His 1 5 10 180 10 PRT Homo sapiens 180 Lys AlaGln Leu Arg Val Gly Tyr Met His 1 5 10 181 10 PRT Homo sapiens 181 LysAla Gln Ser Ser Val Gly Tyr Met His 1 5 10 182 10 PRT Homo sapiens 182Lys Ala Gln Leu Ser Val Gly Tyr Met His 1 5 10 183 10 PRT Homo sapiens183 Lys Ala Ser Ser Arg Val Gly Tyr Met His 1 5 10 184 10 PRT Homosapiens 184 Lys Ala Ser Leu Arg Val Gly Tyr Met His 1 5 10 185 10 PRTHomo sapiens 185 Lys Ala Ser Ser Ser Val Gly Tyr Met His 1 5 10 186 10PRT Homo sapiens 186 Lys Ala Ser Leu Ser Val Gly Tyr Met His 1 5 10 18710 PRT Homo sapiens 187 Ser Ala Ser Leu Arg Val Gly Tyr Met His 1 5 10188 10 PRT Homo sapiens 188 Ser Ala Ser Leu Ser Val Gly Tyr Met His 1 510 189 10 PRT Homo sapiens 189 Ser Ala Gln Ser Arg Val Gly Tyr Met His 15 10 190 10 PRT Homo sapiens 190 Ser Ala Gln Leu Arg Val Gly Tyr Met His1 5 10 191 10 PRT Homo sapiens 191 Ser Ala Gln Ser Ser Val Gly Tyr MetHis 1 5 10 192 10 PRT Homo sapiens 192 Leu Pro Ser Leu Ser Val Gly TyrMet His 1 5 10 193 10 PRT Homo sapiens 193 Leu Pro Ser Ser Ser Val GlyTyr Met His 1 5 10 194 10 PRT Homo sapiens 194 Leu Pro Ser Leu Arg ValGly Tyr Met His 1 5 10 195 10 PRT Homo sapiens 195 Leu Cys Ser Ser ArgVal Gly Tyr Met His 1 5 10 196 10 PRT Homo sapiens 196 Leu Cys Ser LeuSer Val Gly Tyr Met His 1 5 10 197 10 PRT Homo sapiens 197 Leu Cys SerSer Ser Val Gly Tyr Met His 1 5 10 198 10 PRT Homo sapiens 198 Leu CysSer Leu Arg Val Gly Tyr Met His 1 5 10 199 10 PRT Homo sapiens 199 LeuPro Gln Ser Arg Val Gly Tyr Met His 1 5 10 200 10 PRT Homo sapiens 200Leu Pro Gln Leu Ser Val Gly Tyr Met His 1 5 10 201 10 PRT Homo sapiens201 Leu Pro Gln Ser Ser Val Gly Tyr Met His 1 5 10 202 10 PRT Homosapiens 202 Leu Pro Gln Leu Arg Val Gly Tyr Met His 1 5 10 203 10 PRTHomo sapiens 203 Leu Cys Gln Ser Arg Val Gly Tyr Met His 1 5 10 204 10PRT Homo sapiens 204 Leu Cys Gln Leu Ser Val Gly Tyr Met His 1 5 10 20510 PRT Homo sapiens 205 Leu Cys Gln Ser Ser Val Gly Tyr Met His 1 5 10206 10 PRT Homo sapiens 206 Leu Cys Gln Leu Arg Val Gly Tyr Met His 1 510 207 10 PRT Homo sapiens 207 Ser Ala Gln Leu Ser Val Gly Tyr Met His 15 10 208 450 PRT Homo sapiens 208 Gln Val Thr Leu Arg Glu Ser Gly ProAla Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr PheSer Gly Phe Ser Leu Ser Thr Ser 20 25 30 Gly Met Ser Val Gly Trp Ile ArgGln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp AspAsp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile SerLys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn MetAsp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser Met Ile ThrAsn Trp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr ValSer Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala ProSer Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys LeuVal Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp AsnSer Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 LeuGln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr LeuMet Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp ValSer His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp GlyVal Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln TyrAsn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His GlnAsp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser AsnLys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala LysGly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser ArgGlu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val LysGly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn GlyGln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu AspSer Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 LysSer Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440445 Gly Lys 450 209 213 PRT Homo sapiens 209 Asp Ile Gln Met Thr Gln SerPro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile ThrCys Lys Cys Gln Leu Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln LysPro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Lys Leu AlaSer Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu PheThr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr TyrTyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly ThrLys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe IlePhe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser ValVal Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val GlnTrp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe195 200 205 Asn Arg Gly Glu Cys 210 210 450 PRT Homo sapiens 210 Gln ValThr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 ThrLeu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 GlyMet Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 TrpLeu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 LeuLys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Ala Arg Ser Met Ile Thr Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr ValSer 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr PhePro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser ValVal Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile CysAsn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg ValGlu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys ProAla Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe ProPro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu ValThr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val LysPhe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys ThrLys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val SerVal Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 GluTyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro ProVal 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys LeuThr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser CysSer Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys SerLeu Ser Leu Ser Pro 435 440 445 Gly Lys 450 211 213 PRT Homo sapiens 211Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 1015 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 2530 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Asp Thr Phe Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Phe Ser Gly Tyr Pro Phe Thr 8590 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser GlyThr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg GluAla Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly AsnSer Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser ThrTyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr GluLys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu SerSer Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 212 450PRT Homo sapiens 212 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val LysPro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe SerLeu Ser Thr Pro 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro GlyLys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys HisTyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr SerLys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala AspThr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr PheAsp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala SerThr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys SerThr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp TyrPhe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala LeuThr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser GlyLeu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser LeuGly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser AsnThr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys ThrHis Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr TyrArg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu AsnGly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu ProAla Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro ArgGlu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met ThrLys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr ProSer Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu AsnAsn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly SerPhe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp GlnGln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu HisAsn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450213 213 PRT Homo sapiens 213 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr LeuSer Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Leu SerSer Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys AlaPro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Tyr Leu Ser Ser Gly Val ProSer Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr IleSer Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe GlnGly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu IleLys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro SerAsp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu LeuAsn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val AspAsn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr GluGln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu ThrLeu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys GluVal Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 AsnArg Gly Glu Cys 210 214 450 PRT Homo sapiens 214 Gln Val Thr Leu Arg GluSer Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu ThrCys Thr Phe Ser Gly Phe Ser Leu Ser Thr Pro 20 25 30 Gly Met Ser Val GlyTrp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp IleTrp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg LeuThr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys ValThr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg AspMet Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr ThrVal Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe ProLeu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 LeuGly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn HisLys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys SerCys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu LeuLeu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro LysAsp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val ValVal Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp TyrVal Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg GluGlu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr ValLeu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys LysVal Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile SerLys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu ProPro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr CysLeu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 GluSer Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu SerPro 435 440 445 Gly Lys 450 215 213 PRT Homo sapiens 215 Asp Ile Gln MetThr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg ValThr Ile Thr Cys Ser Leu Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp TyrGln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr ArgGly Leu Pro Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser GlyThr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp PheAla Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe GlyGly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 SerVal Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu SerSer 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys ValTyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val ThrLys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 216 450 PRT Homo sapiens216 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 510 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Pro 2025 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 3540 45 Trp Leu Ala Asp Ile Trp Trp Asp Gly Lys Lys His Tyr Asn Pro Ser 5055 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 6570 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro SerVal 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly ThrAla Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val HisThr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu SerSer Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr TyrIle Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp LysArg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro ProCys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe LeuPhe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr ProGlu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro GluVal Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn AlaLys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 ValVal Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val SerLeu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala ValGlu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr SerLys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val PheSer Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr GlnLys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 217 213 PRT Homosapiens 217 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Pro Ser Ser Arg Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Met Arg Leu Ala Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr ValAla Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln LeuLys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu GlnSer Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser LysAsp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys AlaAsp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His GlnGly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys210 218 450 PRT Homo sapiens 218 Gln Val Thr Leu Arg Glu Ser Gly Pro AlaLeu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe SerGly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg GlnPro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp GlyLys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser LysAsp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met AspPro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe AsnTrp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val SerSer Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro SerSer Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu ValLys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn SerGly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu GlnSer Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 SerSer Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu MetIle 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val SerHis Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly ValGlu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr AsnSer Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln AspTrp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn LysAla Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys GlyGln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg GluGlu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys GlyPhe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly GlnPro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp SerAsp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys SerArg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 GluAla Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys 450 219 213 PRT Homo sapiens 219 Asp Ile Gln Met Thr Gln Ser ProSer Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr CysSer Leu Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys ProGly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Ser SerGly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe ThrLeu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr TyrCys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr LysVal Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile PhePro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val ValCys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln TrpLys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 SerVal Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195200 205 Asn Arg Gly Glu Cys 210 220 450 PRT Homo sapiens 220 Gln Val ThrLeu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr LeuThr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly MetSer Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp LeuAla Asp Ile Trp Trp Asp Gly Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu LysAsp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 ValLeu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 CysAla Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe ProAla Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val ValThr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys AsnVal Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val GluPro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro AlaPro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro ProLys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val ThrCys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys PheAsn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr LysPro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser ValLeu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu TyrLys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 LysThr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu ThrVal Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys SerVal Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser LeuSer Leu Ser Pro 435 440 445 Gly Lys 450 221 213 PRT Homo sapiens 221 AspIle Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val Gly Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Phe Lys Leu Ser Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 9095 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu AlaLys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn SerGln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr TyrSer Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu LysHis Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser SerPro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 222 450 PRTHomo sapiens 222 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys ProThr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser LeuSer Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly LysAla Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Gly Lys Lys Ser TyrAsn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser LysAsn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp ThrAla Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe AspVal Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser ThrLys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser ThrSer Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr PhePro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu ThrSer Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly LeuTyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu GlyThr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn ThrLys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr HisThr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 ProSer Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn GlyLys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro AlaPro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg GluPro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr LysAsn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro SerAsp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn AsnTyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser PhePhe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln GlnGly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His AsnHis Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 223213 PRT Homo sapiens 223 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Leu Ser SerArg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Met Tyr Gln Ser Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile LysArg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser AspGlu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu AsnAsn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp AsnAla Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu GlnAsp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr LeuSer Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu ValThr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn ArgGly Glu Cys 210 224 450 PRT Homo sapiens 224 Gln Val Thr Leu Arg Glu SerGly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr CysThr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly TrpIle Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile TrpTrp Asp Gly Lys Lys Ser Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu ThrIle Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val ThrAsn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp MetIle Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr ValThr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro LeuAla Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu GlyCys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser CysAsp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu LeuGly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys AspThr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val ValAsp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr ValAsp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu GluGln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val LeuHis Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys ValSer Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser LysAla Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro ProSer Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys LeuVal Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu SerAsn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro435 440 445 Gly Lys 450 225 213 PRT Homo sapiens 225 Asp Ile Gln Met ThrGln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val ThrIle Thr Cys Leu Pro Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr GlnGln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Met TyrGln Ser Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly ThrGlu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe AlaThr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly GlyGly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser ValPhe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 AlaSer Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val TyrAla 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr LysSer Phe 195 200 205 Asn Arg Gly Glu Cys 210 226 450 PRT Homo sapiens 226Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 1015 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 2530 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 4045 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser 50 5560 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 7075 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 8590 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro SerVal 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly ThrAla Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val HisThr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu SerSer Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr TyrIle Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp LysArg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro ProCys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe LeuPhe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr ProGlu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro GluVal Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn AlaLys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 ValVal Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val SerLeu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala ValGlu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr SerLys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val PheSer Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr GlnLys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 227 213 PRT Homosapiens 227 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Phe Phe Leu Asp Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr ValAla Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln LeuLys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu GlnSer Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser LysAsp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys AlaAsp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His GlnGly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys210 228 450 PRT Homo sapiens 228 Gln Val Thr Leu Arg Glu Ser Gly Pro AlaLeu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe SerGly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg GlnPro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp AspLys Lys Ser Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser LysAsp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met AspPro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe AsnTrp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val SerSer Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro SerSer Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu ValLys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn SerGly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu GlnSer Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 SerSer Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu MetIle 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val SerHis Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly ValGlu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr AsnSer Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln AspTrp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn LysAla Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys GlyGln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg GluGlu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys GlyPhe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly GlnPro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp SerAsp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys SerArg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 GluAla Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys 450 229 213 PRT Homo sapiens 229 Asp Ile Gln Met Thr Gln Ser ProSer Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr CysSer Pro Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys ProGly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Arg Tyr Gln Ser SerGly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe ThrLeu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr TyrCys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr LysVal Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile PhePro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val ValCys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln TrpLys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 SerVal Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195200 205 Asn Arg Gly Glu Cys 210 230 451 PRT Homo sapiens 230 Gln Val ThrLeu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr LeuThr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser 20 25 30 Gly MetSer Val Gly Tyr Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu 35 40 45 Glu TrpLeu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro 50 55 60 Ser LeuLys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln 65 70 75 80 ValVal Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr 85 90 95 TyrCys Ala Arg Ser Met Ile Thr Asn Trp Tyr Phe Asp Val Trp Gly 100 105 110Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr PhePro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser ValVal Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile CysAsn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Arg ValGlu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys ProAla Pro Glu Leu Leu Gly 225 230 235 240 Gly Pro Ser Val Phe Leu Phe ProPro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu ValThr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val LysPhe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys ThrLys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val SerVal Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 305 310 315 320 Lys GluTyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 GluLys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys LeuThr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser CysSer Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys SerLeu Ser Leu Ser 435 440 445 Pro Gly Lys 450 231 213 PRT Homo sapiens 231Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 1015 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 2530 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 4045 Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 5560 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 7075 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 8590 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser GlyThr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg GluAla Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly AsnSer Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser ThrTyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr GluLys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu SerSer Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 232 450PRT Homo sapiens 232 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val LysPro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe SerLeu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro GlyLys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys AspTyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr SerLys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala AspThr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr PheAsp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala SerThr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys SerThr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp TyrPhe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala LeuThr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser GlyLeu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser LeuGly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser AsnThr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys ThrHis Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr TyrArg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu AsnGly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu ProAla Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro ArgGlu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met ThrLys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr ProSer Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu AsnAsn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly SerPhe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp GlnGln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu HisAsn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450233 213 PRT Homo sapiens 233 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr LeuSer Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala SerSer Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys AlaPro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Ala Ser Gly Val ProSer Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr IleSer Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe GlnGly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu IleLys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro SerAsp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu LeuAsn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val AspAsn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr GluGln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu ThrLeu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys GluVal Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 AsnArg Gly Glu Cys 210 234 450 PRT Homo sapiens 234 Gln Val Thr Leu Arg GluSer Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu ThrCys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val GlyTrp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp IleTrp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg LeuThr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys ValThr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg AspMet Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr ThrVal Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe ProLeu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 LeuGly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn HisLys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys SerCys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu LeuLeu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro LysAsp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val ValVal Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp TyrVal Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg GluGlu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr ValLeu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys LysVal Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile SerLys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu ProPro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr CysLeu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 GluSer Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu SerPro 435 440 445 Gly Lys 450 235 213 PRT Homo sapiens 235 Asp Ile Gln MetThr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg ValThr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 25 30 His Trp TyrGln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr TyrLys Gln Thr Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser GlyThr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp PheAla Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe GlyGly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 SerVal Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu SerSer 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys ValTyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val ThrLys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 236 450 PRT Homo sapiens236 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 510 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 2025 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 3540 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 5055 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 6570 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro SerVal 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly ThrAla Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val HisThr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu SerSer Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr TyrIle Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp LysArg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro ProCys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe LeuPhe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr ProGlu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro GluVal Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn AlaLys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 ValVal Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val SerLeu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala ValGlu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr SerLys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val PheSer Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr GlnLys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 237 213 PRT Homosapiens 237 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Arg Tyr Leu Ser Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr ValAla Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln LeuLys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu GlnSer Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser LysAsp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys AlaAsp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His GlnGly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys210 238 450 PRT Homo sapiens 238 Gln Val Thr Leu Arg Glu Ser Gly Pro AlaLeu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe SerGly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg GlnPro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp AspLys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser LysAsp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met AspPro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Thr AsnPhe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val SerSer Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro SerSer Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu ValLys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn SerGly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu GlnSer Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 SerSer Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu MetIle 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val SerHis Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly ValGlu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr AsnSer Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln AspTrp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn LysAla Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys GlyGln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg GluGlu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys GlyPhe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly GlnPro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp SerAsp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys SerArg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 GluAla Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys 450 239 213 PRT Homo sapiens 239 Asp Ile Gln Met Thr Gln Ser ProSer Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr CysSer Ala Ser Ser Ser Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys ProGly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Ala SerGly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe ThrLeu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr TyrCys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr LysVal Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile PhePro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val ValCys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln TrpLys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 SerVal Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195200 205 Asn Arg Gly Glu Cys 210 240 450 PRT Homo sapiens 240 Gln Val ThrLeu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr LeuThr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly MetSer Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp LeuAla Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu LysSer Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 ValLeu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 CysAla Arg Ser Met Ile Thr Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe ProAla Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val ValThr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys AsnVal Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val GluPro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro AlaPro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro ProLys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val ThrCys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys PheAsn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr LysPro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser ValLeu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu TyrLys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 LysThr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu ThrVal Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys SerVal Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser LeuSer Leu Ser Pro 435 440 445 Gly Lys 450 241 213 PRT Homo sapiens 241 AspIle Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val Gly Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Phe Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Phe Tyr Pro Phe Thr 85 9095 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu AlaLys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn SerGln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr TyrSer Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu LysHis Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser SerPro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 242 450 PRTHomo sapiens 242 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys ProThr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser LeuSer Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly LysAla Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp TyrAsn Pro Ser 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser LysAsn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp ThrAla Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Thr Asn Phe Tyr Phe AspVal Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser ThrLys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser ThrSer Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr PhePro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu ThrSer Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly LeuTyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu GlyThr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn ThrLys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr HisThr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 ProSer Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn GlyLys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro AlaPro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg GluPro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr LysAsn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro SerAsp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn AsnTyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser PhePhe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln GlnGly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His AsnHis Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 243213 PRT Homo sapiens 243 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser SerSer Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe Lys Leu Thr Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile LysArg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser AspGlu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu AsnAsn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp AsnAla Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu GlnAsp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr LeuSer Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu ValThr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn ArgGly Glu Cys 210 244 450 PRT Homo sapiens 244 Gln Val Thr Leu Arg Glu SerGly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr CysThr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly TrpIle Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile TrpTrp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Arg Leu ThrIle Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val ThrAsn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp MetIle Thr Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr ValThr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro LeuAla Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu GlyCys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser CysAsp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu LeuGly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys AspThr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val ValAsp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr ValAsp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu GluGln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val LeuHis Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys ValSer Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser LysAla Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro ProSer Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys LeuVal Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu SerAsn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro435 440 445 Gly Lys 450 245 213 PRT Homo sapiens 245 Asp Ile Gln Met ThrGln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val ThrIle Thr Cys Ser Ala Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr GlnGln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Phe LysLeu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly ThrGlu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe AlaThr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly GlyGly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser ValPhe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 AlaSer Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val TyrAla 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr LysSer Phe 195 200 205 Asn Arg Gly Glu Cys 210 246 450 PRT Homo sapiens 246Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 1015 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 2530 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 4045 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys Asp Tyr Asn Pro Ser 50 5560 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 7075 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 8590 95 Cys Ala Arg Asp Met Ile Thr Asn Phe Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro SerVal 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly ThrAla Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val HisThr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu SerSer Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr TyrIle Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp LysArg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro ProCys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe LeuPhe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr ProGlu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro GluVal Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn AlaLys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 ValVal Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val SerLeu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala ValGlu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr SerLys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val PheSer Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr GlnLys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 247 213 PRT Homosapiens 247 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Phe Arg Leu Ala Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr ValAla Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln LeuLys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu GlnSer Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser LysAsp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys AlaAsp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His GlnGly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys210 248 450 PRT Homo sapiens 248 1 5 10 15 Thr Leu Thr Leu Thr Cys ThrPhe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp IleArg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp TrpAsp Asp Lys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr IleSer Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr AsnMet Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ser Met IleThr Asn Trp Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val ThrVal Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu AlaPro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly CysLeu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 TrpAsn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu GlyGly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp ThrLeu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val AspVal Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val AspGly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu GlnTyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu HisGln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val SerAsn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys AlaLys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro SerArg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu ValLys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser AsnGly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 LeuAsp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435440 445 Gly Lys 450 249 214 PRT Homo sapiens 249 Asp Ile Gln Met Thr GlnSer Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr IleThr Cys Ser Pro Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln GlnLys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Tyr Arg HisLeu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly ThrGlu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe AlaThr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe 85 90 95 Thr Phe Gly GlyGly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser ValPhe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr AlaSer Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 LysVal Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr LysSer 195 200 205 Phe Asn Arg Gly Glu Cys 210 250 450 PRT Homo sapiens 250Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 1015 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 2530 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 4045 Trp Leu Ala Asp Ile Trp Trp Asp Gly Lys Lys His Tyr Asn Pro Ser 50 5560 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 7075 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 8590 95 Cys Ala Arg Asp Met Ile Phe Asn Trp Tyr Phe Asp Val Trp Gly Gln100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro SerVal 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly ThrAla Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro ValThr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val HisThr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu SerSer Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr TyrIle Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp LysArg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro ProCys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe LeuPhe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr ProGlu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro GluVal Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn AlaLys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 ValVal Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val SerLeu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala ValGlu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr ThrPro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr SerLys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val PheSer Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr GlnLys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 251 213 PRT Homosapiens 251 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser ValGly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Leu Ser Ser Ser Val GlyTyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu LeuIle Tyr 35 40 45 Asp Thr Phe Phe His Arg Ser Gly Val Pro Ser Arg Phe SerGly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu GlnPro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly TyrPro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr ValAla Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln LeuLys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe TyrPro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu GlnSer Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser LysAsp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys AlaAsp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His GlnGly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys210 252 450 PRT Homo sapiens 252 Gln Val Thr Leu Arg Glu Ser Gly Pro AlaLeu Val Lys Pro Thr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe SerGly Phe Ser Leu Ser Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg GlnPro Pro Gly Lys Ala Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp AspLys Lys His Tyr Asn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser LysAsp Thr Ser Lys Asn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met AspPro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe AsnPhe Tyr Phe Asp Val Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val SerSer Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro SerSer Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu ValLys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn SerGly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu GlnSer Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 SerSer Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu MetIle 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val SerHis Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly ValGlu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr AsnSer Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln AspTrp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn LysAla Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys GlyGln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg GluGlu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys GlyPhe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly GlnPro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp SerAsp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys SerArg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 GluAla Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys 450 253 213 PRT Homo sapiens 253 Asp Ile Gln Met Thr Gln Ser ProSer Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr CysSer Ala Ser Ser Arg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys ProGly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Asp Thr Leu Leu Leu Asp SerGly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe ThrLeu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr TyrCys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr LysVal Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile PhePro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val ValCys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln TrpLys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 SerVal Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195200 205 Asn Arg Gly Glu Cys 210 254 450 PRT Homo sapiens 254 Gln Val ThrLeu Arg Glu Ser Gly Pro Ala Leu Val Lys Pro Thr Gln 1 5 10 15 Thr LeuThr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ala 20 25 30 Gly MetSer Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45 Trp LeuAla Asp Ile Trp Trp Asp Asp Lys Lys His Tyr Asn Pro Ser 50 55 60 Leu LysAsp Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 ValLeu Lys Val Thr Asn Met Asp Pro Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 CysAla Arg Asp Met Ile Phe Asn Phe Tyr Phe Asp Val Trp Gly Gln 100 105 110Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe ProAla Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val ValThr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys AsnVal Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Arg Val GluPro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro AlaPro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro ProLys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val ThrCys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys PheAsn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr LysPro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser ValLeu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu TyrLys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 LysThr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu ThrVal Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys SerVal Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser LeuSer Leu Ser Pro 435 440 445 Gly Lys 450 255 213 PRT Homo sapiens 255 AspIle Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Ser Arg Val Gly Tyr Met 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Asp 65 70 7580 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly Ser Gly Tyr Pro Phe Thr 85 9095 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu AlaLys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn SerGln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr TyrSer Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu LysHis Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser SerPro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 256 450 PRTHomo sapiens 256 Gln Val Thr Leu Arg Glu Ser Gly Pro Ala Leu Val Lys ProThr Gln 1 5 10 15 Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser LeuSer Thr Ala 20 25 30 Gly Met Ser Val Gly Trp Ile Arg Gln Pro Pro Gly LysAla Leu Glu 35 40 45 Trp Leu Ala Asp Ile Trp Trp Asp Asp Lys Lys His TyrAsn Pro Ser 50 55 60 Leu Lys Asp Arg Leu Thr Ile Ser Lys Asp Thr Ser LysAsn Gln Val 65 70 75 80 Val Leu Lys Val Thr Asn Met Asp Pro Ala Asp ThrAla Thr Tyr Tyr 85 90 95 Cys Ala Arg Asp Met Ile Phe Asn Phe Tyr Phe AspVal Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser ThrLys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser ThrSer Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr PhePro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu ThrSer Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly LeuTyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu GlyThr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn ThrLys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr HisThr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 ProSer Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn GlyLys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro AlaPro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg GluPro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr LysAsn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro SerAsp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn AsnTyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser PhePhe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln GlnGly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His AsnHis Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 257213 PRT Homo sapiens 257 Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu SerAla Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser SerArg Val Gly Tyr Met 20 25 30 His Trp Tyr Gln Gln Lys Pro Gly Lys Ala ProLys Leu Leu Ile Tyr 35 40 45 Asp Thr Ser Phe Leu Asp Ser Gly Val Pro SerArg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile SerSer Leu Gln Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Phe Gln GlySer Gly Tyr Pro Phe Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile LysArg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser AspGlu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu AsnAsn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp AsnAla Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu GlnAsp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr LeuSer Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu ValThr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn ArgGly Glu Cys 210 258 26 DNA Artificial Sequence Description of ArtificialSequence Primer 258 agtgtcttaa ccagcaaagt gttaga 26 259 26 DNAArtificial Sequence Description of Artificial Sequence Primer 259tcattgactt gagatattga tgcatc 26

What is claimed is:
 1. A method of preventing treating or amelioratingone or more symptoms associated with a RSV infection in a mammal, saidmethod comprising administering to said mammal a dose of an effectiveamount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens, wherein saideffective amount is less than 15 mg/kg of said antibodies or antibodyfragments.
 2. The method of claim 2, wherein said antibodies or antibodyfragments have an affinity of at least 2×10⁸ M⁻¹ for said one or moreRSV antigens.
 3. The method of claim 2 or 3, wherein the dose is lessthan 5 mg/kg or less.
 4. The method of claim 2 or 3, wherein the dose is3 mg/kg or less, or 1.5 mg/kg or less.
 5. The method of claim 2, whereinsaid antibodies or antibody fragments are administered by a nebulizer orinhaler.
 6. The method of claim 2, wherein said antibodies or antibodyfragments are administered intramuscularly, intravaneously orsubcutaneously.
 7. The method of claim 2, wherein said antibodies orantibody fragments administered 1, 2, 3, 4 or 5 times during the RSVseason.
 8. The method of claim 2, wherein at least one of the antibodiesis a human or humanized monoclonal antibody.
 9. The method of claim 2,wherein the mammal is a human subject, a human subject which has had abone marrow transplant, an elderly human subject, or a human subjectwhich has cystic fibrosis, bronchopulmonary dysplasia, congenital heartdisease, congenital immunodeficiency or acquired immunodeficiency. 10.The method of claim 2, wherein the mammal is a human infant.
 11. Themethod of claim 2, wherein the mammal is a human infant born prematurelyor is at risk of hospitalization for a RSV infection.
 12. The method ofclaim 2, wherein at least one of the antibodies is SYNAGIS®, AFFF,P12f2, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR,H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5,A4B4(1), A4B4-L1FR-S28R, or A4B4-F52S.
 13. A method of preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, comprising administering to said mammal a firstdose of an effective amount of one or more antibodies thatimmunospecifically bind to one or more RSV antigens, wherein saideffective amount is a dose of less than 15 mg/kg of said antibodies orantibody fragments, wherein said administration results in an effectiveserum titer of said antibodies or antibody fragments that is less than30 μg/ml at least 20 days after the administration of said first doseand prior to the administration of a subsequent dose.
 14. The method ofclaim 13, wherein said antibodies or antibody fragments bind to said oneor more RSV antigens with an affinity constant of at least 2×10⁸ M⁻¹.15. The method of claim 13 or 14, wherein the dose is less than 5 mg/kgor less.
 16. The method of claim 13 or 14, wherein the dose is 3 mg/kgor less, or 1.5 mg/kg or less.
 17. The method of claim 14, wherein saideffective serum titer is at least 2 μg/ml.
 18. The method of claim 14,wherein said effective serum titer is less than 30 μg/ml at least 30days after the administration of said first dose and prior to theadministration of a subsequent dose.
 19. The method of claim 14, whereinthe dose is 1.5 mg/kg or less and said effective serum titer is at least2 μg/ml at least 30 days after the administration of said first dose andprior to the administration of a subsequent dose.
 20. The method ofclaim 14, wherein said antibodies or antibody fragments are administeredby a nebulizer or inhaler.
 21. The method of claim 14, wherein saidantibodies or antibody fragments are administered intramuscularly,intravaneously or subcutaneously.
 22. The method of claim 14, whereinsaid antibodies or antibody fragments have half-lives in said humansubject of greater than 25 days.
 23. The method of claim 14, wherein atleast one of the antibodies is a human or humanized monoclonal antibody.24. The method of claim 14, wherein the mammal is a human subject, ahuman subject which has had a bone marrow transplant, an elderly humansubject, or a human subject which has cystic fibrosis, bronchopulmonarydysplasia, congenital heart disease, congenital immunodeficiency oracquired immunodeficiency.
 25. The method of claim 14, wherein themammal is a human infant.
 26. The method of claim 14, wherein the mammalis a human infant born prematurely or is at risk of hospitalization fora RSV infection.
 27. The method of claim 14, wherein at least one of theantibodies is SYNAGIS®, AFFF, P12f2, P12f4, P11d4, A1e9, A12a6, A13c4,A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8,L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4-L1FR-S28R, or A4B4-F52S.28. A method of preventing, treating or ameliorating one or moresymptoms associated with a RSV infection in a mammal, said methodcomprising administering to said mammal a first dose of an effectiveamount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens, wherein saideffective amount is approximately 15 mg/kg or less of said antibodies orantibody fragments and an effective serum titer is maintained for atleast 20 days after the administration said first dose and prior to theadministration of a subsequent dose.
 29. The method of claim 28, whereinthe antibodies or antibody fragments have an affinity of at least 2×10⁸M¹⁻ for said one or more RSV antigens.
 30. The method of claim 28,wherein said effective serum titer is at least 30 μg/ml of saidantibodies or antibody fragments.
 31. The method of claim 28, whereinsaid effective serum titer is at least 2 μg/ml of said antibodies orantibody fragments.
 32. The method of claim 28, wherein the effectiveserum titer is maintained for at least 25 days or at least 30 days. 33.The method of claim 28, wherein said antibodies or antibody fragmentsare administered by a nebulizer or inhaler.
 34. The method of claim 28,wherein said antibodies or antibody fragments are administeredintramuscularly, intravaneously or subcutaneously.
 35. The method ofclaim 28, wherein said antibodies or antibody fragments have half-livesin said human subject of greater than 25 days.
 36. The method of claim28, wherein at least one of the antibodies is a human or humanizedmonoclonal antibody.
 37. The method of claim 28, wherein the mammal is ahuman subject, a human subject which has had a bone marrow transplant,an elderly human subject, or a human subject which has cystic fibrosis,bronchopulmonary dysplasia, congenital heart disease, congenitalimmunodeficiency or acquired immunodeficiency.
 38. The method of claim28, wherein the mammal is a human infant.
 39. The method of claim 28,wherein the mammal is a human infant born prematurely or is at risk ofhospitalization for a RSV infection.
 40. The method of claim 28, whereinat least one of the antibodies is SYNAGIS®, AFFF, P12f2, P12f4, P11d4,A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6,DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4-L1FR-S28R, or A4B4-F52S.
 41. A sustained release formulationcomprising one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens.
 42. Apharmaceutical composition comprising one or more antibodies orfragments thereof that immunospecifically bind to one or more RSVantigens formulated for pulmonary delivery.
 43. The sustained releaseformulation of claim 41, wherein the antibodies or antibody fragmentshave an affinity of at least 2×10⁸ M¹⁻ for said one or more RSVantigens.
 44. The pharmaceutical composition of claim 42, wherein theantibodies or antibody fragments have an affinity of at least 2×10⁸ M¹⁻for said one or more RSV antigens.
 45. The sustained release formulationof claim 41, wherein at least one of the antibodies or antibodyfragments is SYNAGIS® or an antigen-binding fragment thereof.
 46. Thepharmaceutical composition of claim 42, wherein at least one of theantibodies or antibody fragments is SYNAGIS® or an antigen-bindingfragment thereof.
 47. The sustained release formulation of claim 41,wherein at least one of said antibodies or antibody fragments is a humanor humanized antibody or antibody fragment.
 48. The pharmaceuticalcomposition of claim 42, wherein at least one of said antibodies orantibody fragments is a human or humanized antibody or antibodyfragment.
 49. The sustained release formulation of claim 41, wherein atleast one of said antibodies is SYNAGIS®, AFFF, P12f2, P12f4, P 11d4,A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6,DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4-L1FR-S28R, or A4B4-F52S.
 50. The pharmaceutical composition ofclaim 42, wherein at least one of said antibodies at least one of theantibodies is SYNAGIS®, AFFF, P12f2, P12f4, P 11d4, A1e9, A12a6, A13c4,A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8,L1-7E5, L2-15B10, A13a11, A1h5, A4B4(1), A4B4-L1FR-S28R, or A4B4-F52S.51. The sustained release formulation of claim 41, wherein at least oneof said antibodies or antibody fragments has an increased in vivohalf-life.
 52. The pharmaceutical composition of claim 42, wherein atleast one of said antibodies or antibody fragments has an increased invivo half-life.
 53. A method of preventing, treating or ameliorating oneor more symptoms associated with a RSV infection in a mammal, saidmethod comprising administering to said mammal an effective amount ofthe sustained release formulation of claim
 41. 54. A method ofpreventing, treating or ameliorating one or more symptoms associatedwith a RSV infection in a mammal, said method comprising administeringto the lungs of said mammal an effective amount of the pharmaceuticalcomposition of claim
 42. 55. The method of claim 53, wherein thesustained release formulation is administered intramuscularly,intravaneously or subcutaneously.
 56. The method of claim 53, whereinthe sustained release formulation is administered by a nebulizer orinhaler.
 57. The method of claim 54, wherein the pharmaceuticalcomposition is administered by a nebulizer or inhaler.
 58. The method ofclaim 53, wherein the mammal is a human subject.
 59. The method of claim54, wherein the mammal is a human subject.
 60. The method of claim 58,wherein the human subject has had a bone marrow transplant, is elderly,or has cystic fibrosis, bronchopulmonary dysplasia, congenital heartdisease, congenital immunodeficiency or acquired immunodeficiency. 61.The method of claim 59, wherein the human subject has had a bone marrowtransplant, is elderly, or has cystic fibrosis, bronchopulmonarydysplasia, congenital heart disease, congenital immunodeficiency oracquired immunodeficiency.
 62. The method of claim 58, wherein the humansubject is an infant.
 63. The method of claim 58, wherein the humansubject is an infant born prematurely or is at risk of hospitalizationfor a RSV infection.
 64. The method of claim 59, wherein the humansubject is an infant.
 65. The method of claim 59, wherein the humansubject is an infant born prematurely or is at risk of hospitalizationfor a RSV infection.
 66. A method of preventing, treating orameliorating one or more symptoms associated with a RSV infection in amammal, said method comprising administering to said mammal a first doseof an effective dose of SYNAGIS® or an antigen-binding fragment thereofin a sustained release formulation, wherein said effective dose isapproximately 15 mg/kg or less of SYNAGIS® or an antigen-bindingfragment thereof and an effective serum titer of at least 30 μg/ml ismaintained for at least 20 days after the administration said first doseand prior to the administration of a subsequent dose.
 67. The method ofclaim 66, wherein said effective serum titer is maintained for at least25 days or at least 30 days after the administration of the first doseand prior to the administration of a subsequent dose.
 68. The method ofclaim 66, wherein SYNAGIS® or an antigen-binding fragment thereof isadministered by a nebulizer or inhaler.
 69. The method of claim 66,wherein SYNAGIS® or an antigen-binding fragment thereof is administeredintramuscularly, intravaneously or subcutaneously.
 70. The method ofclaim 66, wherein SYNAGIS® or an antigen-binding fragment thereof isadministered 1, 2, 3, 4, or 5 times during the RSV season.
 71. Themethod of claim 66, wherein the mammal is a human subject, a humansubject which has had a bone marrow transplant, an elderly humansubject, or a human subject which has cystic fibrosis, bronchopulmonarydysplasia, congenital heart disease, congenital immunodeficiency oracquired immunodeficiency.
 72. The method of claim 66, wherein themammal is a human infant.
 73. The method of claim 66, wherein the mammalis a human infant born prematurely or is at risk of hospitalization fora RSV infection.
 74. A method of preventing, treating or amelioratingone or more symptoms associated with a RSV infection in a mammal, saidmethod comprising administering to said mammal a first dose of aneffective dose of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens with an affinity ofat least 2×10⁸ M¹⁻ in a sustained release formulation, wherein saideffective dose is approximately 15 mg/kg or less of said antibodies orantibody fragments and an effective serum titer of less than 30 μg/ml ismaintained for at least 20 days after the administration said first doseand prior to the administration of a subsequent dose.
 75. The method ofclaim 74, wherein said effective serum titer is at least 2 μg/ml. 76.The method of claim 74, wherein said effective serum titer is maintainedfor at least 25 days or at least 30 days after the administration of thefirst dose and prior to the administration of a subsequent dose.
 77. Themethod of claim 74, wherein said antibodies or antibody fragments areadministered by a nebulizer or inhaler.
 78. The method of claim 74,wherein said antibodies or antibody fragments are administeredintramuscularly, intravaneously or subcutaneously.
 79. The method ofclaim 74, wherein said antibodies or antibody fragments are administered1, 2, 3, 4, or 5 times during the RSV season.
 80. The method of claim74, wherein said antibodies or antibody fragments have half-lives insaid human subject of greater than 25 days.
 81. The method of claim 74,wherein at least one of the antibodies is a human or humanizedmonoclonal antibody.
 82. The method of claim 74, wherein the mammal is ahuman subject, a human subject which has had a bone marrow transplant,an elderly human subject, or a human subject which has cystic fibrosis,bronchopulmonary dysplasia, congenital heart disease, congenitalimmunodeficiency or acquired immunodeficiency.
 83. The method of claim74, wherein the mammal is a human infant.
 84. The method of claim 74,wherein the mammal is a human infant born prematurely or is at risk ofhospitalization for a RSV infection. .
 85. The method of claim 74,wherein at least one of the antibodies is AFFF, P12f2, P12f4, P11d4,A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR, H3-3F4, M3H9, Y10H6,DG, AFFF(1), 6H8, L1 -7E5, L2-15B10, A13a11, A1h5, A4B4(1),A4B4-L1FR-S28R, or A4B4-F52S.
 86. A method of preventing, treating orameliorating one or more symptoms associated with a RSV infection in amammal, said method comprising administering to said mammal a dose of aneffective amount of one or more antibodies or fragments thereof thatimmunospecifically bind to one or more RSV antigens and have increasedin vivo half-lives, wherein said effective amount is a doseapproximately 15 mg/kg or less of said antibodies or antibody fragments.87. The method of claim 86, wherein said antibodies or antibodyfragments have an affinity of at least 2×10 M⁻¹ for said one or more RSVantigens.
 88. The method of claim 86, wherein the dose is less than 5mg/kg or less, 3 mg/kg or less, or 1.5 mg/kg or less.
 89. The method ofclaim 86, wherein the increase in in vivo half-life is from 21 days toat least 25 days or from 21 days to at least 30 days.
 90. A method ofpreventing, treating or ameliorating one or more symptoms associatedwith a RSV infection in a mammal, said method comprising administeringto said mammal a dose of an effective amount of HL-SYNAGIS or anantigen-binding fragment thereof, wherein said effective amount is adose of approximately 15 mg/kg or less of HL-SYNAGIS® or anantigen-binding fragment thereof which results in an effective serumtiter that is at least 30 μg/ml at least 30 days after theadministration of said first dose and prior to the administration of asubsequent dose.
 91. The method of claim 86 or 90, wherein saidantibodies or antibody fragments are administered by a nebulizer orinhaler.
 92. The method of claim 86 or 90, wherein said antibodies orantibody fragments are administered intramuscularly, intravaneously orsubcutaneously.
 93. The method of claim 86 or 90, wherein the mammal isa human subject, a human subject which has had a bone marrow transplant,an elderly human subject, or a human subject which has cystic fibrosis,bronchopulmonary dysplasia, congenital heart disease, congenitalimmunodeficiency or acquired immunodeficiency.
 94. The method of claim86 or 90, wherein the mammal is a human infant.
 95. The method of claim86 or 90, wherein the mammal is a human infant born prematurely or is atrisk of hospitalization for a RSV infection.
 96. A method of preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, said method comprising administering to saidmammal a dose of an effective amount of one or more antibodies orfragments thereof, wherein said antibodies or fragments thereofimmunospecifically bind to one or more RSV antigens and have increasedin vivo half-lives, and wherein said effective amount is a dose ofapproximately 15 mg/kg or less of said antibodies or antibody fragmentswhich results in an effective serum titer of less than 30 μg/ml at least30 days after the administration of said first dose and prior to theadministration of a subsequent dose.
 97. The method of claim 96, whereinsaid antibodies or antibody fragments have an affinity of at least 2×10⁸M⁻¹ for said one or more RSV antigens.
 98. The method of claim 96,wherein the effective serum titer is at least 2 μg/ml, at least 40μg/ml, or at least 50 μg/ml.
 99. The method of claim 96, wherein theeffective serum titer is at least 30 μg/ml at least 35 days after theadministration of said first dose and prior to the administration of asubsequent dose.
 100. The method of claim 96, wherein the effectiveserum titer is at least 2 μg/ml at least 35 days after theadministration of said first dose and prior to the administration of asubsequent dose.
 101. The method of claim 90, wherein HL-SYNAGIS or anantigen-binding fragment thereof is formulated in a sustained releaseformulation.
 102. The method of claim 86 or 96, wherein said antibodiesor fragments thereof are formulated in a sustained release formulation.103. The method of claim 96, wherein said antibodies or fragmentsthereof are administered by a nebulizer or inhaler.
 104. The method ofclaim 96, wherein said antibodies or fragments thereof are administeredintramuscularly, intravaneously or subcutaneously.
 105. The method ofclaim 90, wherein HL-SYNAGIS or an antigen-binding fragment thereof hasa half-life in said mammalian subject of greater than 25 days.
 106. Themethod of claim 96, wherein said antibodies or fragments thereof havehalf-lives in said mammalian subject of greater than 25 days.
 107. Themethod of claim 86 or 96, wherein at least one of the antibodies is ahuman or humanized monoclonal antibody.
 108. The method of claim 96,wherein the mammal is a human subject, a human subject which has had abone marrow transplant, an elderly human subject, or a human subjectwhich has cystic fibrosis, bronchopulmonary dysplasia, congenital heartdisease, congenital immunodeficiency or acquired immunodeficiency. 109.The method of claim 96, wherein the mammal is a human infant.
 110. Themethod of claim 96, wherein the mammal is a human infant bornprematurely or is at risk of hospitalization for a RSV infection. 111.The method of claim 86 or 96, wherein at least one of said antibodiescomprises a VH CDR1 having the amino acid sequence of SEQ ID NO: 1, SEQID NO: 10, SEQ ID NO: 18, a VH CDR2 having the amino acid sequence ofSEQ ID NO: 2, SEQ ID NO: 19, SEQ ID NO: 25, SEQ ID NO: 37, SEQ ID NO:41, SEQ ID NO: 45, SEQ ID NO: 82, SEQ ID NO: 86, SEQ ID NO: 91, SEQ IDNO: 93, SEQ ID NO: 100, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 109,SEQ ID NO: 111, or SEQ ID NO: 1 14, a VH CDR3 having the amino acidsequence of SEQ ID NO: 3, SEQ ID NO: 12, SEQ ID NO: 20, SEQ ID NO: 29,SEQ ID NO: 79, SEQ ID NO: 83, SEQ ID NO: 94 or SEQ ID NO: 97, a VL CDR1having the amino acid sequence of SEQ ID NO: 4, SEQ ID NO: 14, SEQ IDNO: 22, SEQ ID NO: 31, SEQ ID NO: 39 or SEQ ID NO: 47, SEQ ID NO: 80,SEQ ID NO: 84, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 92, SEQ ID NO:95, SEQ ID NO: 98, SEQ ID NO: 101, SEQ ID NO: 104, SEQ ID NO: 107, SEQID NO: 1 10, SEQ ID NO: 1 12, SEQ ID NO: 1 15, SEQ ID NO: 117, SEQ IDNO: 119, SEQ ID NO: 120, SEQ ID NO: 122, SEQ ID NO: 125, SEQ ID NO: 127,SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 132, SEQ ID NO: 134, SEQ IDNO: 136, SEQ ID NO: 138, SEQ ID NO: 140, SEQ ID NO: 142, SEQ ID NO: 144,SEQ ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ IDNO: 153, SEQ ID NO: 155, SEQ ID NO: 157, SEQ ID NO: 159, SEQ ID NO: 161,SEQ ID NO: 163, SEQ ID NO: 166, SEQ ID NO: 168, SEQ ID NO: 169, SEQ IDNO: 171, SEQ ID NO: 173, SEQ ID NO: 175, SEQ ID NO: 177, SEQ ID NO: 179,SEQ ID NO: 180, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ IDNO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 188,SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, SEQ ID NO: 192, SEQ IDNO: 193, SEQ ID NO: 194, SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197,SEQ ID NO: 198, SEQ ID NO: 199, SEQ ID NO: 200, SEQ ID NO: 201, SEQ IDNO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205, SEQ ID NO: 206,or SEQ ID NO: 207, a VL CDR2 having the amino acid sequence of SEQ IDNO: 5, SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 27, SEQ ID NO: 32, SEQID NO: 35, SEQ ID NO: 43, SEQ ID NO: 50, SEQ ID NO: 53, SEQ ID NO: 57,SEQ ID NO: 59, SEQ ID NO: 63, SEQ ID NO: 66, SEQ ID NO: 69, SEQ ID NO:73, SEQ ID NO: 75, SEQ ID NO: 77, SEQ ID NO: 81, SEQ ID NO: 85, SEQ IDNO: 88, SEQ ID NO: 90, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 102, SEQID NO: 105, SEQ ID NO: 108, SEQ ID NO: 113, SEQ ID NO: 116, SEQ ID NO:118, SEQ ID NO: 121, SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 126, SEQID NO: 128, SEQ ID NO: 131, SEQ ID NO: 133, SEQ ID NO: 135, SEQ ID NO:137, SEQ ID NO: 139, SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, SEQID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 154, SEQ ID NO:156, SEQ ID NO: 158, SEQ ID NO: 160, SEQ ID NO: 162, SEQ ID NO: 164, SEQID NO: 165, SEQ ID NO: 167, SEQ ID NO: 170, SEQ ID NO: 172, SEQ ID NO:174, SEQ ID NO: 176, or SEQ ID NO: 178, or a VL CDR3 having the aminoacid sequence of SEQ ID NO: 6, SEQ ID NO: 16, or SEQ ID NO:
 61. 112. Amethod of preventing, treating or ameliorating one or more symptomsassociated with a RSV infection in a mammal, said method comprisingadministering to the lungs of said mammal a first dose of an effectiveamount of a composition comprising one or more antibodies or fragmentsthereof that immunospecifically bind to one or more RSV antigens,wherein said effective amount results in an effective concentration ofat least 20 ng per mg of lung protein at least 20 days after theadministration said first dose and prior to the administration of asubsequent dose.
 113. The method of claim 112, wherein said antibodiesor antibody fragments have an affinity of at least 2×10⁸ M⁻¹ for saidone or more RSV antigens.
 114. The method of claim 112, wherein saidantibodies or antibody fragments have in vivo half-lives of greater than30 days.
 115. The method of claim 112, wherein said antibodies orantibody fragments are administered by a nebulizer or inhaler.
 116. Themethod of claim 112, wherein said antibodies or antibody fragments areadministered intramuscularly, intravaneously or subcutaneously.
 117. Themethod of claim 112, wherein at least one of said antibodies is a humanor humanized monoclonal antibody.
 118. The method of claim 112, whereinthe mammal is a human subject, a human subject which has had a bonemarrow transplant, an elderly human subject, or a human subject whichhas cystic fibrosis, bronchopulmonary dysplasia, congenital heartdisease, congenital immunodeficiency or acquired immunodeficiency. 119.The method of claim 112, wherein the mammal is a human infant.
 120. Themethod of claim 112, wherein the mammal is a human infant bornprematurely or is at risk of hospitalization for a RSV infection. 121.The method of claim 112, wherein at least one of the antibodies is AFFF,P12f2, P12f4, P11d4, A1e9, A12a6, A13c4, A17d4, A4B4, A8C7, 1X-493L1FR,H3-3F4, M3H9, Y10H6, DG, AFFF(1), 6H8, L1-7E5, L2-15B10, A13a11, A1h5,A4B4(1), A4B4-L1FR-S28R, or A4B4-F52S.
 122. A method of preventing,treating or ameliorating one or more symptoms associated with a RSVinfection in a mammal, said method comprising administering to the lungsof said mammal a first dose of an effective amount of a compositioncomprising SYNAGIS® or a fragment thereof, wherein said effective amountresults in an effective concentration of at least 20 ng per mg of lungprotein at least 20 days after the administration said first dose andprior to the administration of a subsequent dose.
 123. The method ofclaim 122, wherein SYNAGIS® or an antigen-binding fragment thereof isadministered by a nebulizer or inhaler.
 124. The method of claim 122,wherein SYNAGIS® or an antigen-binding fragment thereof is administeredintramuscularly, intravaneously or subcutaneously.
 125. The method ofclaim 122, wherein the mammal is a human subject, a human subject whichhas had a bone marrow transplant, an elderly human subject, or a humansubject which has cystic fibrosis, bronchopulmonary dysplasia,congenital heart disease, congenital immunodeficiency or acquiredimmunodeficiency.
 126. The method of claim 122, wherein the mammal is ahuman infant.
 127. The method of claim 122, wherein the mammal is ahuman infant born prematurely or is at risk of hospitalization for a RSVinfection.